expression vector pet15b  (Millipore)


Bioz Verified Symbol Millipore is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92

    Structured Review

    Millipore expression vector pet15b
    Lysis assays with cells of C. difficile strain 11204. Cells were grown to mid-exponential phase, harvested by centrifugation, and then either flash-frozen in liquid nitrogen and stored at −20°C before being assayed (A) or used immediately (B to D). Cells were incubated with Ni-NTA column-purified endolysin (E1) from E. coli expressing CD27L from <t>pET15b-</t> cd27l (A to C) or with crude protein extracts (D). (A) Effect of CD27L (7 μg, black squares, or 0.7 μg, gray squares) on frozen cells compared to that of the buffer control (EB; x's). (B) Effects of different amounts of CD27L (10.5 μg, black rectangles; 3.5 μg, gray rectangles; 0.7 μg, white rectangles; 0.35 μg, black squares; and 0.07 μg, gray squares) on fresh cells compared to that of the buffer control (EB; x's). (C) Activity profile of CD27L (black symbols) compared to that of the buffer control (EB; white symbols) tested at pH 4.5 (black squares), 5.8 (gray squares), 6.5 (black diamonds), 7.0 (gray diamonds), 7.6 (black triangles), and 8.3 (gray triangles). (D) Lysis of cells incubated with 50 μg of crude protein extracts from E. coli expressing pET15b- cd27l (black triangles) or pET15b (white triangles) or from L. lactis expressing pUK200His- cd27l (black diamonds) or pUK200His (white diamonds) or with the TN buffer control (plus signs). Values are the means of results from duplicate assays ± standard deviations.
    Expression Vector Pet15b, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/expression vector pet15b/product/Millipore
    Average 92 stars, based on 110 article reviews
    Price from $9.99 to $1999.99
    expression vector pet15b - by Bioz Stars, 2020-04
    92/100 stars

    Images

    1) Product Images from "Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin ▿ Bacteriophage and Its Cloned Biologically Active Endolysin ▿ †"

    Article Title: Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin ▿ Bacteriophage and Its Cloned Biologically Active Endolysin ▿ †

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.00686-08

    Lysis assays with cells of C. difficile strain 11204. Cells were grown to mid-exponential phase, harvested by centrifugation, and then either flash-frozen in liquid nitrogen and stored at −20°C before being assayed (A) or used immediately (B to D). Cells were incubated with Ni-NTA column-purified endolysin (E1) from E. coli expressing CD27L from pET15b- cd27l (A to C) or with crude protein extracts (D). (A) Effect of CD27L (7 μg, black squares, or 0.7 μg, gray squares) on frozen cells compared to that of the buffer control (EB; x's). (B) Effects of different amounts of CD27L (10.5 μg, black rectangles; 3.5 μg, gray rectangles; 0.7 μg, white rectangles; 0.35 μg, black squares; and 0.07 μg, gray squares) on fresh cells compared to that of the buffer control (EB; x's). (C) Activity profile of CD27L (black symbols) compared to that of the buffer control (EB; white symbols) tested at pH 4.5 (black squares), 5.8 (gray squares), 6.5 (black diamonds), 7.0 (gray diamonds), 7.6 (black triangles), and 8.3 (gray triangles). (D) Lysis of cells incubated with 50 μg of crude protein extracts from E. coli expressing pET15b- cd27l (black triangles) or pET15b (white triangles) or from L. lactis expressing pUK200His- cd27l (black diamonds) or pUK200His (white diamonds) or with the TN buffer control (plus signs). Values are the means of results from duplicate assays ± standard deviations.
    Figure Legend Snippet: Lysis assays with cells of C. difficile strain 11204. Cells were grown to mid-exponential phase, harvested by centrifugation, and then either flash-frozen in liquid nitrogen and stored at −20°C before being assayed (A) or used immediately (B to D). Cells were incubated with Ni-NTA column-purified endolysin (E1) from E. coli expressing CD27L from pET15b- cd27l (A to C) or with crude protein extracts (D). (A) Effect of CD27L (7 μg, black squares, or 0.7 μg, gray squares) on frozen cells compared to that of the buffer control (EB; x's). (B) Effects of different amounts of CD27L (10.5 μg, black rectangles; 3.5 μg, gray rectangles; 0.7 μg, white rectangles; 0.35 μg, black squares; and 0.07 μg, gray squares) on fresh cells compared to that of the buffer control (EB; x's). (C) Activity profile of CD27L (black symbols) compared to that of the buffer control (EB; white symbols) tested at pH 4.5 (black squares), 5.8 (gray squares), 6.5 (black diamonds), 7.0 (gray diamonds), 7.6 (black triangles), and 8.3 (gray triangles). (D) Lysis of cells incubated with 50 μg of crude protein extracts from E. coli expressing pET15b- cd27l (black triangles) or pET15b (white triangles) or from L. lactis expressing pUK200His- cd27l (black diamonds) or pUK200His (white diamonds) or with the TN buffer control (plus signs). Values are the means of results from duplicate assays ± standard deviations.

    Techniques Used: Lysis, Centrifugation, Incubation, Purification, Expressing, Activity Assay

    (A) Gel analysis of crude protein lysates from E. coli and L. lactis expressing ΦCD27 endolysin and empty vector controls. Lanes: 1, SeeBlue marker (Invitrogen); 2 to 5, lysates from L. lactis containing pUK200His- cd27l (2 and 3) or pUK200His (4 and 5) with (2 and 4) or without (3 and 5) nisin induction; and 6 and 7, lysates from E. coli containing pET15b- cd27l (6) or pET15b (7), both with IPTG induction. Arrows indicate the His-tagged endolysin—when it is expressed from pET15b- cd27l with the His tag MGSSHHHHHHSSGLVPRGSH, the size is ca. 32 kDa, and when it is expressed from pUK200His- cd27l with the His tag MSHHHHHHA, the size is ca. 31 kDa. (B) Western analysis of the gel in panel A with a six-His tag antibody. (C) Gel analysis of Ni-NTA column-purified His-tagged endolysin from E. coli expressing pET15b- cd27l . Lanes: 1, SeeBlue marker; 2 to 5, E. coli (pET15b- cd27l ) total protein extracts after 5 h of induction with IPTG; 2, crude lysate; 3, column flowthrough; 4, primary wash effluent; 5, secondary wash effluent; 6, primary eluate (E1); and 7, secondary eluate (E2). Crude protein lysates were loaded at 10 μg of total protein per lane, while lanes loaded with samples from Ni-NTA column fractionation contained 6.5 μl per lane.
    Figure Legend Snippet: (A) Gel analysis of crude protein lysates from E. coli and L. lactis expressing ΦCD27 endolysin and empty vector controls. Lanes: 1, SeeBlue marker (Invitrogen); 2 to 5, lysates from L. lactis containing pUK200His- cd27l (2 and 3) or pUK200His (4 and 5) with (2 and 4) or without (3 and 5) nisin induction; and 6 and 7, lysates from E. coli containing pET15b- cd27l (6) or pET15b (7), both with IPTG induction. Arrows indicate the His-tagged endolysin—when it is expressed from pET15b- cd27l with the His tag MGSSHHHHHHSSGLVPRGSH, the size is ca. 32 kDa, and when it is expressed from pUK200His- cd27l with the His tag MSHHHHHHA, the size is ca. 31 kDa. (B) Western analysis of the gel in panel A with a six-His tag antibody. (C) Gel analysis of Ni-NTA column-purified His-tagged endolysin from E. coli expressing pET15b- cd27l . Lanes: 1, SeeBlue marker; 2 to 5, E. coli (pET15b- cd27l ) total protein extracts after 5 h of induction with IPTG; 2, crude lysate; 3, column flowthrough; 4, primary wash effluent; 5, secondary wash effluent; 6, primary eluate (E1); and 7, secondary eluate (E2). Crude protein lysates were loaded at 10 μg of total protein per lane, while lanes loaded with samples from Ni-NTA column fractionation contained 6.5 μl per lane.

    Techniques Used: Expressing, Plasmid Preparation, Marker, Western Blot, Purification, Fractionation

    2) Product Images from "Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed"

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0118075

    In vivo assay for cAMP phosphodiesterase acivity of PdeM. (A) β-Galactosidase activity of E coli BL21 (DE3) harbouring pET15b or its derivative pET-PdeM without or with induction by IPTG. (B) SDS-PAGE of the total proteins extracted from E coli BL21 (DE3) harbouring pET15b or its derivative pET-PdeM without or with induction by 0.5 mM IPTG. Lane M: protein molecular weight marker; lane pET: induced cells of E coli BL21 (DE3) harbouring pET15b, and lane pET-PdeM: induced cells of E . coli BL21 (DE3) harbouring pET-PdeM.
    Figure Legend Snippet: In vivo assay for cAMP phosphodiesterase acivity of PdeM. (A) β-Galactosidase activity of E coli BL21 (DE3) harbouring pET15b or its derivative pET-PdeM without or with induction by IPTG. (B) SDS-PAGE of the total proteins extracted from E coli BL21 (DE3) harbouring pET15b or its derivative pET-PdeM without or with induction by 0.5 mM IPTG. Lane M: protein molecular weight marker; lane pET: induced cells of E coli BL21 (DE3) harbouring pET15b, and lane pET-PdeM: induced cells of E . coli BL21 (DE3) harbouring pET-PdeM.

    Techniques Used: In Vivo, Activity Assay, Positron Emission Tomography, SDS Page, Molecular Weight, Marker

    3) Product Images from "Molecular Characterization of a Novel Glucosyltransferase from Lactobacillus reuteri Strain 121 Synthesizing a Unique, Highly Branched Glucan with ?-(1- > 4) and ?-(1- > 6) Glucosidic Bonds"

    Article Title: Molecular Characterization of a Novel Glucosyltransferase from Lactobacillus reuteri Strain 121 Synthesizing a Unique, Highly Branched Glucan with ?-(1- > 4) and ?-(1- > 6) Glucosidic Bonds

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.68.9.4283-4291.2002

    Overview of primers and restriction sites used for cloning of the gtfA gene in the expression vector pET15B (Novagen).
    Figure Legend Snippet: Overview of primers and restriction sites used for cloning of the gtfA gene in the expression vector pET15B (Novagen).

    Techniques Used: Clone Assay, Expressing, Plasmid Preparation

    4) Product Images from "Sublingual Immunization with M2-Based Vaccine Induces Broad Protective Immunity against Influenza"

    Article Title: Sublingual Immunization with M2-Based Vaccine Induces Broad Protective Immunity against Influenza

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0027953

    Construction of plasmids and purification of M2 proteins. (A) The synthetic M2eC or 3M2eC genes without hydrophobic region (amino acids 26–55) from PR8 virus were cloned into pET15b vector (B). The recombinant proteins expressed in E. coli were purified by His-tag affinity chromatography and detected by Western blot using M2e-specific monoclonal Ab, 14C2.
    Figure Legend Snippet: Construction of plasmids and purification of M2 proteins. (A) The synthetic M2eC or 3M2eC genes without hydrophobic region (amino acids 26–55) from PR8 virus were cloned into pET15b vector (B). The recombinant proteins expressed in E. coli were purified by His-tag affinity chromatography and detected by Western blot using M2e-specific monoclonal Ab, 14C2.

    Techniques Used: Purification, Clone Assay, Plasmid Preparation, Recombinant, Affinity Chromatography, Western Blot

    5) Product Images from "Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed"

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0118075

    In vivo assay for cAMP phosphodiesterase acivity of PdeM. (A) β-Galactosidase activity of E coli BL21 (DE3) harbouring pET15b or its derivative pET-PdeM without or with induction by IPTG. (B) SDS-PAGE of the total proteins extracted from E coli BL21 (DE3) harbouring pET15b or its derivative pET-PdeM without or with induction by 0.5 mM IPTG. Lane M: protein molecular weight marker; lane pET: induced cells of E coli BL21 (DE3) harbouring pET15b, and lane pET-PdeM: induced cells of E . coli BL21 (DE3) harbouring pET-PdeM.
    Figure Legend Snippet: In vivo assay for cAMP phosphodiesterase acivity of PdeM. (A) β-Galactosidase activity of E coli BL21 (DE3) harbouring pET15b or its derivative pET-PdeM without or with induction by IPTG. (B) SDS-PAGE of the total proteins extracted from E coli BL21 (DE3) harbouring pET15b or its derivative pET-PdeM without or with induction by 0.5 mM IPTG. Lane M: protein molecular weight marker; lane pET: induced cells of E coli BL21 (DE3) harbouring pET15b, and lane pET-PdeM: induced cells of E . coli BL21 (DE3) harbouring pET-PdeM.

    Techniques Used: In Vivo, Activity Assay, Positron Emission Tomography, SDS Page, Molecular Weight, Marker

    Related Articles

    Clone Assay:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: Paragraph title: Cloning and expression of EqCXCL16 in Escherichia coli . ... The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes.

    Article Title: Structural Insight into the Recognition of r(UAG) by Musashi-1 RBD2, and Construction of a Model of Musashi-1 RBD1-2 Bound to the Minimum Target RNA
    Article Snippet: .. Protein and RNA Preparation The RBD2+, comprising mouse Msi1 residues Lys109-Arg200, containing RBD2, was cloned into the expression vector pET15b (Novagen), as a fusion with an N-terminal hexahistidine affinity tag and thrombin cleavage site. ..

    Article Title: Molecular Characterization of a Novel Glucosyltransferase from Lactobacillus reuteri Strain 121 Synthesizing a Unique, Highly Branched Glucan with ?-(1- > 4) and ?-(1- > 6) Glucosidic Bonds
    Article Snippet: The resulting fragments (2,547 and 2,723 bp) were cloned in the corresponding sites of pBluescript II SK(+), yielding pBSP2500 and pBPA2700, respectively. .. Plasmid pBGTF1 was digested with Nco I/ Bam HI, and the resulting 5.3-kb fragment was ligated into the corresponding sites of the expression vector pET15b (Novagen) yielding p15gtf.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: Paragraph title: Cloning, overexpression and purification of metagenomic phosphodiesterase ... The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM.

    Article Title: Identification of functionally relevant histidine residues in the apoptotic nuclease CAD
    Article Snippet: Paragraph title: cDNA cloning ... The coding region for hICAD-L (DFF45) was inserted in-frame as an Nco I– Sal I fragment into the Nco I and Xho I sites of the bacterial expression vector pET15b (Novagen).

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. Cloning, overexpression and purification of metagenomic phosphodiesterase A construct for heterologous expression of the gene encoding metagenomic phosphodiesterase (PdeM) was constructed by PCR amplification from the fosmid clone and cloning into the expression vector pET15b (Novagen). .. The complete coding region of the PdeM gene was amplified by PCR using primers pdemF/pdemR designed from the annotated gene sequence (Froward 5’-GGGAATTC CATATG AGCCCCCAAGCTTCACCG-3’ and Reverse 5’-CGC GGATCC TCAAGTTCCCGGCAAGGCCC-3’).

    Centrifugation:

    Article Title: Crystallization and preliminary X-ray analysis of a dye-linked d-lactate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix
    Article Snippet: The amplified 1.4 kbp fragment was digested with Nde I and Bam HI and then ligated with the expression vector pET15b (Novagen, Madison, Wisconsin, USA) previously linearized using Nde I and Bam HI, yielding pDLDH. .. Expression was then induced by adding 0.1 m M isopropyl β- d -1-thiogalactopyranoside to the medium and cultivation was continued for an additional 21 h at 293 K. The cells were then harvested by centrifugation, suspended in 50 m M sodium phosphate buffer pH 8.0 supplemented with 300 m M NaCl (buffer A ) and disrupted by ultrasonication.

    Article Title: Characterization of Drosophila Tyramine β-HydroxylaseGene and Isolation of Mutant Flies Lacking Octopamine
    Article Snippet: The 1.2 kb Sal – Xho fragment of the DmDBH cDNA (Fig. B ) was inserted into the Xho I site of the bacterial expression vector pET15b (Novagen, Madison, WI) in frame with the His6 tag and under the bacteriophage T7 promoter present in the vector. .. After 30 min centrifugation at 10,000 rpm, the lysate was adsorbed to a nickel chelate affinity resin (Qiagen, Chatsworth, CA) column, and the protein was eluted with a pH gradient in the above buffer.

    Amplification:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The amplified EqCXCL16 fragment (amino acids [aa] 17 to 247) was run on a 1% agarose gel (E-Gel EX; Life Technologies, Grand Island, NY) and purified with a commercial kit (Zymoclean gel recovery kit; Zymo Research, Irvine, CA). .. The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes.

    Article Title: Amino Acids Important for DNA Recognition by the Response Regulator OmpR *Amino Acids Important for DNA Recognition by the Response Regulator OmpR * S⃞
    Article Snippet: Construction of ompR Mutations by PCR —To create the OmpR mutants, ompR was amplified using OmpR061 and OmpR062 or EnvZ061 primers and then subcloned into the pGEM-T easy vector. .. Overexpression and Purification of OmpR Mutants —To overproduce the mutant OmpR proteins, each plasmid was cleaved with NdeI and BamHI and ligated into the expression vector pET15b (Novagen) that had been cleaved with the same enzymes.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM. .. E . coli DH5α was then transformed with the ligation mix and the transformants were selected on Luria agar with ampicillin (100 μg mL-1 ).

    Article Title: Crystallization and preliminary X-ray analysis of a dye-linked d-lactate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix
    Article Snippet: .. The amplified 1.4 kbp fragment was digested with Nde I and Bam HI and then ligated with the expression vector pET15b (Novagen, Madison, Wisconsin, USA) previously linearized using Nde I and Bam HI, yielding pDLDH. .. E. coli strain Rosetta-gami 2 (DE3) (Stratagene, La Jolla, California, USA) was then transformed with the vector and the transformants were cultivated at 310 K in 1 l SB medium (1.2% tryptone peptone, 2.4% yeast extract, 1.25% K2 HPO4 , 0.38% KH2 PO4 and 0.5% glycerol) containing 50 µg ml−1 ampicillin and 50 µg ml−1 chloramphenicol until the optical density at 600 nm reached 0.6.

    Article Title: Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin ▿ Bacteriophage and Its Cloned Biologically Active Endolysin ▿ †
    Article Snippet: The ΦCD27 endolysin gene cd27l was amplified from genomic DNA by using primers designed to create an NdeI site at the 5′ end (5′-TTA CAT ATG AAA ATA TGT ATA ACA GTA GG, where underlining indicates the bases corresponding to the restriction endonuclease site) and a XhoI site downstream of the coding sequence (5′-CAA CCA C CT CGA G TT GAT AAC). .. NdeI- and XhoI-restricted PCR products were ligated into the expression vector pET15b (Novagen) by using Fast-Link DNA ligase (Epicentre), and the resulting construct, pET15b- cd27l , was introduced into chemically competent E. coli TOP10 (Invitrogen).

    Article Title: Identification of functionally relevant histidine residues in the apoptotic nuclease CAD
    Article Snippet: To isolate the cDNA for human ICAD-L/DFF45 (hICAD-L), total RNA was extracted from HeLa cells with a RNeasy Mini Kit (Qiagen) and used for first-strand cDNA synthesis and subsequent amplification of the hICAD-L gene with the Access RT–PCR system (Promega). .. The coding region for hICAD-L (DFF45) was inserted in-frame as an Nco I– Sal I fragment into the Nco I and Xho I sites of the bacterial expression vector pET15b (Novagen).

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. Cloning, overexpression and purification of metagenomic phosphodiesterase A construct for heterologous expression of the gene encoding metagenomic phosphodiesterase (PdeM) was constructed by PCR amplification from the fosmid clone and cloning into the expression vector pET15b (Novagen). .. The complete coding region of the PdeM gene was amplified by PCR using primers pdemF/pdemR designed from the annotated gene sequence (Froward 5’-GGGAATTC CATATG AGCCCCCAAGCTTCACCG-3’ and Reverse 5’-CGC GGATCC TCAAGTTCCCGGCAAGGCCC-3’).

    DNA Ligation:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: .. The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes. .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Synthesized:

    Article Title: Sublingual Immunization with M2-Based Vaccine Induces Broad Protective Immunity against Influenza
    Article Snippet: Construction of plasmids expressing M2eC or 3M2eC protein A gene ( ) encoding three tandem copies of M2e conjugated to C-terminus sequence of M2 protein without residues 26–55 from influenza A/Puerto Rico/8/34 (H1N1) virus was chemically synthesized by Bio S & T Inc. (Canada). .. For the plasmid expressing 3M2eC, the gene was digested with Xho I and BamH I and inserted into the bacterial expression vector pET15b (Novagen, Madison, WI) to express as a fusion of his-tag at the N terminus, resulting in the plasmid pET15b-3M2eC.

    Article Title: Molecular Characterization of a Novel Glucosyltransferase from Lactobacillus reuteri Strain 121 Synthesizing a Unique, Highly Branched Glucan with ?-(1- > 4) and ?-(1- > 6) Glucosidic Bonds
    Article Snippet: The product of the first PCR, synthesized with primers GTFpr11 (5′-GATGCAT GAGCTC CCATGG ACCAACAAGTTCAGCAAGCTTCC-3′, containing Sac I [in boldface type] and Nco I [underlined] sites) and GTFpr12 (5′-GTGCATTAAAGTACGTAACCAATCAGTATTTCCGG-3′), was digested with Sac I/ Pst I. .. Plasmid pBGTF1 was digested with Nco I/ Bam HI, and the resulting 5.3-kb fragment was ligated into the corresponding sites of the expression vector pET15b (Novagen) yielding p15gtf.

    Article Title: Characterization of Drosophila Tyramine β-HydroxylaseGene and Isolation of Mutant Flies Lacking Octopamine
    Article Snippet: In situ detection of RNA in whole-mount larval CNS was as described in using digoxygenin-labeled DNA probe synthesized for the Pst – Xho fragment of the cDNA clone (Fig. B ) according to the manufacturer’s protocols (Boehringer Mannheim, Indianapolis, IN). .. The 1.2 kb Sal – Xho fragment of the DmDBH cDNA (Fig. B ) was inserted into the Xho I site of the bacterial expression vector pET15b (Novagen, Madison, WI) in frame with the His6 tag and under the bacteriophage T7 promoter present in the vector.

    Construct:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes. .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Article Title: Sublingual Immunization with M2-Based Vaccine Induces Broad Protective Immunity against Influenza
    Article Snippet: For the plasmid expressing 3M2eC, the gene was digested with Xho I and BamH I and inserted into the bacterial expression vector pET15b (Novagen, Madison, WI) to express as a fusion of his-tag at the N terminus, resulting in the plasmid pET15b-3M2eC. .. For the construct expressing M2eC protein, which has one M2e domain, the synthesized gene was digested with Nde I and BamH I, and then inserted into pET15b vector.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: Cloning, overexpression and purification of metagenomic phosphodiesterase A construct for heterologous expression of the gene encoding metagenomic phosphodiesterase (PdeM) was constructed by PCR amplification from the fosmid clone and cloning into the expression vector pET15b (Novagen). .. The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM.

    Article Title: Crystallization and preliminary X-ray analysis of a dye-linked d-lactate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix
    Article Snippet: To construct the expression plasmid for the putative A. pernix DLDH homologue, a 1.4 kbp gene fragment composed of the gene encoding the enzyme (APE_0487) plus Nde I and Bam HI restriction sites was amplified by PCR with the following two primers. .. The amplified 1.4 kbp fragment was digested with Nde I and Bam HI and then ligated with the expression vector pET15b (Novagen, Madison, Wisconsin, USA) previously linearized using Nde I and Bam HI, yielding pDLDH.

    Article Title: Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin ▿ Bacteriophage and Its Cloned Biologically Active Endolysin ▿ †
    Article Snippet: .. NdeI- and XhoI-restricted PCR products were ligated into the expression vector pET15b (Novagen) by using Fast-Link DNA ligase (Epicentre), and the resulting construct, pET15b- cd27l , was introduced into chemically competent E. coli TOP10 (Invitrogen). .. Positive transformants were selected with ampicillin (100 μg/ml), gene identity was confirmed by sequencing, and chemically competent E. coli BL21(DE3) (Invitrogen) was transformed with the plasmid constructs or the original vector pET15b for protein expression.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. Cloning, overexpression and purification of metagenomic phosphodiesterase A construct for heterologous expression of the gene encoding metagenomic phosphodiesterase (PdeM) was constructed by PCR amplification from the fosmid clone and cloning into the expression vector pET15b (Novagen). .. The complete coding region of the PdeM gene was amplified by PCR using primers pdemF/pdemR designed from the annotated gene sequence (Froward 5’-GGGAATTC CATATG AGCCCCCAAGCTTCACCG-3’ and Reverse 5’-CGC GGATCC TCAAGTTCCCGGCAAGGCCC-3’).

    Incubation:

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM. .. For overexpression and purification of recombinant protein, the recombinant E . coli BL21 (DE3) cells grown overnight in LB medium containing ampicillin (100 μg mL-1 ) at 37°C were inoculated in fresh LB medium (1:100) containing ampicillin (100 μg mL-1 ) and incubated at 37°C with shaking at 170 rpm.

    Activity Assay:

    Article Title: Crystallization and preliminary X-ray analysis of a dye-linked d-lactate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix
    Article Snippet: The amplified 1.4 kbp fragment was digested with Nde I and Bam HI and then ligated with the expression vector pET15b (Novagen, Madison, Wisconsin, USA) previously linearized using Nde I and Bam HI, yielding pDLDH. .. DLDH activity in the crude extract was assayed as described previously (Satomura et al. , 2008 ).

    Expressing:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: .. The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes. .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Article Title: Sublingual Immunization with M2-Based Vaccine Induces Broad Protective Immunity against Influenza
    Article Snippet: .. For the plasmid expressing 3M2eC, the gene was digested with Xho I and BamH I and inserted into the bacterial expression vector pET15b (Novagen, Madison, WI) to express as a fusion of his-tag at the N terminus, resulting in the plasmid pET15b-3M2eC. .. For the construct expressing M2eC protein, which has one M2e domain, the synthesized gene was digested with Nde I and BamH I, and then inserted into pET15b vector.

    Article Title: Structural Insight into the Recognition of r(UAG) by Musashi-1 RBD2, and Construction of a Model of Musashi-1 RBD1-2 Bound to the Minimum Target RNA
    Article Snippet: .. Protein and RNA Preparation The RBD2+, comprising mouse Msi1 residues Lys109-Arg200, containing RBD2, was cloned into the expression vector pET15b (Novagen), as a fusion with an N-terminal hexahistidine affinity tag and thrombin cleavage site. ..

    Article Title: Amino Acids Important for DNA Recognition by the Response Regulator OmpR *Amino Acids Important for DNA Recognition by the Response Regulator OmpR * S⃞
    Article Snippet: .. Overexpression and Purification of OmpR Mutants —To overproduce the mutant OmpR proteins, each plasmid was cleaved with NdeI and BamHI and ligated into the expression vector pET15b (Novagen) that had been cleaved with the same enzymes. .. The ligation mixture was used to transform E. coli strain DH5α, and individual transformants were screened for the presence of the appropriate recombinant plasmid.

    Article Title: Molecular Characterization of a Novel Glucosyltransferase from Lactobacillus reuteri Strain 121 Synthesizing a Unique, Highly Branched Glucan with ?-(1- > 4) and ?-(1- > 6) Glucosidic Bonds
    Article Snippet: .. Plasmid pBGTF1 was digested with Nco I/ Bam HI, and the resulting 5.3-kb fragment was ligated into the corresponding sites of the expression vector pET15b (Novagen) yielding p15gtf. ..

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM. .. E . coli DH5α was then transformed with the ligation mix and the transformants were selected on Luria agar with ampicillin (100 μg mL-1 ).

    Article Title: Crystallization and preliminary X-ray analysis of a dye-linked d-lactate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix
    Article Snippet: .. The amplified 1.4 kbp fragment was digested with Nde I and Bam HI and then ligated with the expression vector pET15b (Novagen, Madison, Wisconsin, USA) previously linearized using Nde I and Bam HI, yielding pDLDH. .. E. coli strain Rosetta-gami 2 (DE3) (Stratagene, La Jolla, California, USA) was then transformed with the vector and the transformants were cultivated at 310 K in 1 l SB medium (1.2% tryptone peptone, 2.4% yeast extract, 1.25% K2 HPO4 , 0.38% KH2 PO4 and 0.5% glycerol) containing 50 µg ml−1 ampicillin and 50 µg ml−1 chloramphenicol until the optical density at 600 nm reached 0.6.

    Article Title: Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin ▿ Bacteriophage and Its Cloned Biologically Active Endolysin ▿ †
    Article Snippet: .. NdeI- and XhoI-restricted PCR products were ligated into the expression vector pET15b (Novagen) by using Fast-Link DNA ligase (Epicentre), and the resulting construct, pET15b- cd27l , was introduced into chemically competent E. coli TOP10 (Invitrogen). .. Positive transformants were selected with ampicillin (100 μg/ml), gene identity was confirmed by sequencing, and chemically competent E. coli BL21(DE3) (Invitrogen) was transformed with the plasmid constructs or the original vector pET15b for protein expression.

    Article Title: Identification of functionally relevant histidine residues in the apoptotic nuclease CAD
    Article Snippet: .. The coding region for hICAD-L (DFF45) was inserted in-frame as an Nco I– Sal I fragment into the Nco I and Xho I sites of the bacterial expression vector pET15b (Novagen). ..

    Article Title: Characterization of Drosophila Tyramine β-HydroxylaseGene and Isolation of Mutant Flies Lacking Octopamine
    Article Snippet: .. The 1.2 kb Sal – Xho fragment of the DmDBH cDNA (Fig. B ) was inserted into the Xho I site of the bacterial expression vector pET15b (Novagen, Madison, WI) in frame with the His6 tag and under the bacteriophage T7 promoter present in the vector. .. The resulting plasmid was introduced into the Escherichia coli BL21(DE3) strain ( ); cultures of the transformed cells were grown to OD600 = 0.5, induced with 1 m m isopropyl β- d -thiogalactopyranoside, then grown for 3 more hr and harvested.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. Cloning, overexpression and purification of metagenomic phosphodiesterase A construct for heterologous expression of the gene encoding metagenomic phosphodiesterase (PdeM) was constructed by PCR amplification from the fosmid clone and cloning into the expression vector pET15b (Novagen). .. The complete coding region of the PdeM gene was amplified by PCR using primers pdemF/pdemR designed from the annotated gene sequence (Froward 5’-GGGAATTC CATATG AGCCCCCAAGCTTCACCG-3’ and Reverse 5’-CGC GGATCC TCAAGTTCCCGGCAAGGCCC-3’).

    Modification:

    Article Title: Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin ▿ Bacteriophage and Its Cloned Biologically Active Endolysin ▿ †
    Article Snippet: NdeI- and XhoI-restricted PCR products were ligated into the expression vector pET15b (Novagen) by using Fast-Link DNA ligase (Epicentre), and the resulting construct, pET15b- cd27l , was introduced into chemically competent E. coli TOP10 (Invitrogen). .. For expression in L. lactis , the cd27l coding sequence was subcloned into the vector pUK200 , which had been modified to include a sequence encoding a six-histidine tag, yielding pUK200His (N. Horn, unpublished data).

    Transformation Assay:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes. .. In order to express recombinant protein, the plasmid was transformed into E. coli BL21(DE3) cells (EMD Millipore Novagen, Temecula, CA).

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM. .. E . coli DH5α was then transformed with the ligation mix and the transformants were selected on Luria agar with ampicillin (100 μg mL-1 ).

    Article Title: Crystallization and preliminary X-ray analysis of a dye-linked d-lactate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix
    Article Snippet: The amplified 1.4 kbp fragment was digested with Nde I and Bam HI and then ligated with the expression vector pET15b (Novagen, Madison, Wisconsin, USA) previously linearized using Nde I and Bam HI, yielding pDLDH. .. E. coli strain Rosetta-gami 2 (DE3) (Stratagene, La Jolla, California, USA) was then transformed with the vector and the transformants were cultivated at 310 K in 1 l SB medium (1.2% tryptone peptone, 2.4% yeast extract, 1.25% K2 HPO4 , 0.38% KH2 PO4 and 0.5% glycerol) containing 50 µg ml−1 ampicillin and 50 µg ml−1 chloramphenicol until the optical density at 600 nm reached 0.6.

    Article Title: Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin ▿ Bacteriophage and Its Cloned Biologically Active Endolysin ▿ †
    Article Snippet: NdeI- and XhoI-restricted PCR products were ligated into the expression vector pET15b (Novagen) by using Fast-Link DNA ligase (Epicentre), and the resulting construct, pET15b- cd27l , was introduced into chemically competent E. coli TOP10 (Invitrogen). .. Positive transformants were selected with ampicillin (100 μg/ml), gene identity was confirmed by sequencing, and chemically competent E. coli BL21(DE3) (Invitrogen) was transformed with the plasmid constructs or the original vector pET15b for protein expression.

    Article Title: Characterization of Drosophila Tyramine β-HydroxylaseGene and Isolation of Mutant Flies Lacking Octopamine
    Article Snippet: The 1.2 kb Sal – Xho fragment of the DmDBH cDNA (Fig. B ) was inserted into the Xho I site of the bacterial expression vector pET15b (Novagen, Madison, WI) in frame with the His6 tag and under the bacteriophage T7 promoter present in the vector. .. The resulting plasmid was introduced into the Escherichia coli BL21(DE3) strain ( ); cultures of the transformed cells were grown to OD600 = 0.5, induced with 1 m m isopropyl β- d -thiogalactopyranoside, then grown for 3 more hr and harvested.

    Over Expression:

    Article Title: Amino Acids Important for DNA Recognition by the Response Regulator OmpR *Amino Acids Important for DNA Recognition by the Response Regulator OmpR * S⃞
    Article Snippet: .. Overexpression and Purification of OmpR Mutants —To overproduce the mutant OmpR proteins, each plasmid was cleaved with NdeI and BamHI and ligated into the expression vector pET15b (Novagen) that had been cleaved with the same enzymes. .. The ligation mixture was used to transform E. coli strain DH5α, and individual transformants were screened for the presence of the appropriate recombinant plasmid.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: Paragraph title: Cloning, overexpression and purification of metagenomic phosphodiesterase ... The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. Cloning, overexpression and purification of metagenomic phosphodiesterase A construct for heterologous expression of the gene encoding metagenomic phosphodiesterase (PdeM) was constructed by PCR amplification from the fosmid clone and cloning into the expression vector pET15b (Novagen). .. The complete coding region of the PdeM gene was amplified by PCR using primers pdemF/pdemR designed from the annotated gene sequence (Froward 5’-GGGAATTC CATATG AGCCCCCAAGCTTCACCG-3’ and Reverse 5’-CGC GGATCC TCAAGTTCCCGGCAAGGCCC-3’).

    Ligation:

    Article Title: Amino Acids Important for DNA Recognition by the Response Regulator OmpR *Amino Acids Important for DNA Recognition by the Response Regulator OmpR * S⃞
    Article Snippet: Overexpression and Purification of OmpR Mutants —To overproduce the mutant OmpR proteins, each plasmid was cleaved with NdeI and BamHI and ligated into the expression vector pET15b (Novagen) that had been cleaved with the same enzymes. .. The ligation mixture was used to transform E. coli strain DH5α, and individual transformants were screened for the presence of the appropriate recombinant plasmid.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM. .. E . coli DH5α was then transformed with the ligation mix and the transformants were selected on Luria agar with ampicillin (100 μg mL-1 ).

    RNA In Situ Hybridization:

    Article Title: Characterization of Drosophila Tyramine β-HydroxylaseGene and Isolation of Mutant Flies Lacking Octopamine
    Article Snippet: Subsequently, the filter was hybridized with a random-primed DNA probe of pRP49 ( ) as a control. .. The 1.2 kb Sal – Xho fragment of the DmDBH cDNA (Fig. B ) was inserted into the Xho I site of the bacterial expression vector pET15b (Novagen, Madison, WI) in frame with the His6 tag and under the bacteriophage T7 promoter present in the vector.

    Introduce:

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: These primers introduce restriction sites for NdeI/BamHI (underline). .. The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM.

    Polymerase Chain Reaction:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The cDNA amplification was carried out according to a standard laboratory PCR protocol using forward primer cx16-15F (5′-GCG CTCGAG GCGTTGCTGACTCTGCAAGG-3′) and reverse primer cx16-15R (5′-GC GGATCC GCACTGCCACTGTAACTGAT-3′) (IDT, Coralville, IA). .. The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes.

    Article Title: Amino Acids Important for DNA Recognition by the Response Regulator OmpR *Amino Acids Important for DNA Recognition by the Response Regulator OmpR * S⃞
    Article Snippet: The product of the second PCR was subcloned into the pGEM-T easy vector. .. Overexpression and Purification of OmpR Mutants —To overproduce the mutant OmpR proteins, each plasmid was cleaved with NdeI and BamHI and ligated into the expression vector pET15b (Novagen) that had been cleaved with the same enzymes.

    Article Title: Molecular Characterization of a Novel Glucosyltransferase from Lactobacillus reuteri Strain 121 Synthesizing a Unique, Highly Branched Glucan with ?-(1- > 4) and ?-(1- > 6) Glucosidic Bonds
    Article Snippet: The product of the second PCR, primed with oligonucleotides GTFpr13 (5′-TTGATGGTATGGTGGCCTAATACTCTTACCC) and GTFpr14 (5′-ATATCGAT GGGCCC C GGATCC TATTA GTGATGGTGATGGTGATG TAGTTTATTTTGATCAAGCATCTTACC-3′, containing Apa I [in boldface type] and Bam HI [underlined] sites and a six-His tag [in italics]), was digested with Pst I/ Apa I. .. Plasmid pBGTF1 was digested with Nco I/ Bam HI, and the resulting 5.3-kb fragment was ligated into the corresponding sites of the expression vector pET15b (Novagen) yielding p15gtf.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM. .. E . coli DH5α was then transformed with the ligation mix and the transformants were selected on Luria agar with ampicillin (100 μg mL-1 ).

    Article Title: Crystallization and preliminary X-ray analysis of a dye-linked d-lactate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix
    Article Snippet: To construct the expression plasmid for the putative A. pernix DLDH homologue, a 1.4 kbp gene fragment composed of the gene encoding the enzyme (APE_0487) plus Nde I and Bam HI restriction sites was amplified by PCR with the following two primers. .. The amplified 1.4 kbp fragment was digested with Nde I and Bam HI and then ligated with the expression vector pET15b (Novagen, Madison, Wisconsin, USA) previously linearized using Nde I and Bam HI, yielding pDLDH.

    Article Title: Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin ▿ Bacteriophage and Its Cloned Biologically Active Endolysin ▿ †
    Article Snippet: .. NdeI- and XhoI-restricted PCR products were ligated into the expression vector pET15b (Novagen) by using Fast-Link DNA ligase (Epicentre), and the resulting construct, pET15b- cd27l , was introduced into chemically competent E. coli TOP10 (Invitrogen). .. Positive transformants were selected with ampicillin (100 μg/ml), gene identity was confirmed by sequencing, and chemically competent E. coli BL21(DE3) (Invitrogen) was transformed with the plasmid constructs or the original vector pET15b for protein expression.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. Cloning, overexpression and purification of metagenomic phosphodiesterase A construct for heterologous expression of the gene encoding metagenomic phosphodiesterase (PdeM) was constructed by PCR amplification from the fosmid clone and cloning into the expression vector pET15b (Novagen). .. The complete coding region of the PdeM gene was amplified by PCR using primers pdemF/pdemR designed from the annotated gene sequence (Froward 5’-GGGAATTC CATATG AGCCCCCAAGCTTCACCG-3’ and Reverse 5’-CGC GGATCC TCAAGTTCCCGGCAAGGCCC-3’).

    Isolation:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes. .. Recombinant plasmid p15-16A (aa 17 to 247) was isolated from ampicillin-resistant clones using a ZR plasmid miniprep kit (Zymo Research, Irvine, CA).

    Sequencing:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The sequence encoding the entire EqCXCL16 without the first 16 amino acid residues from the signal sequences was amplified from cDNA made from mRNA extracted from equine monocytes using a Smart cDNA synthesis kit (Clontech Laboratories Inc., Mountain View, CA). .. The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes.

    Article Title: Sublingual Immunization with M2-Based Vaccine Induces Broad Protective Immunity against Influenza
    Article Snippet: Construction of plasmids expressing M2eC or 3M2eC protein A gene ( ) encoding three tandem copies of M2e conjugated to C-terminus sequence of M2 protein without residues 26–55 from influenza A/Puerto Rico/8/34 (H1N1) virus was chemically synthesized by Bio S & T Inc. (Canada). .. For the plasmid expressing 3M2eC, the gene was digested with Xho I and BamH I and inserted into the bacterial expression vector pET15b (Novagen, Madison, WI) to express as a fusion of his-tag at the N terminus, resulting in the plasmid pET15b-3M2eC.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: The complete coding region of the PdeM gene was amplified by PCR using primers pdemF/pdemR designed from the annotated gene sequence (Froward 5’-GGGAATTC CATATG AGCCCCCAAGCTTCACCG-3’ and Reverse 5’-CGC GGATCC TCAAGTTCCCGGCAAGGCCC-3’). .. The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM.

    Article Title: Crystallization and preliminary X-ray analysis of a dye-linked d-lactate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix
    Article Snippet: The primer 5′-AACATATGGCTCGTATAGCTGAGGAGCTTG-3′ was designed to contain the N-terminal region of the DLDH gene homologue and an Nde I digestion sequence, while the primer 5′-­TTGGATCCTCACTCAGCCGCTACAACCTTC-3′ was designed to contain the C-terminal region and a Bam HI digestion sequence. .. The amplified 1.4 kbp fragment was digested with Nde I and Bam HI and then ligated with the expression vector pET15b (Novagen, Madison, Wisconsin, USA) previously linearized using Nde I and Bam HI, yielding pDLDH.

    Article Title: Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin ▿ Bacteriophage and Its Cloned Biologically Active Endolysin ▿ †
    Article Snippet: The ΦCD27 endolysin gene cd27l was amplified from genomic DNA by using primers designed to create an NdeI site at the 5′ end (5′-TTA CAT ATG AAA ATA TGT ATA ACA GTA GG, where underlining indicates the bases corresponding to the restriction endonuclease site) and a XhoI site downstream of the coding sequence (5′-CAA CCA C CT CGA G TT GAT AAC). .. NdeI- and XhoI-restricted PCR products were ligated into the expression vector pET15b (Novagen) by using Fast-Link DNA ligase (Epicentre), and the resulting construct, pET15b- cd27l , was introduced into chemically competent E. coli TOP10 (Invitrogen).

    Sonication:

    Article Title: Characterization of Drosophila Tyramine β-HydroxylaseGene and Isolation of Mutant Flies Lacking Octopamine
    Article Snippet: The 1.2 kb Sal – Xho fragment of the DmDBH cDNA (Fig. B ) was inserted into the Xho I site of the bacterial expression vector pET15b (Novagen, Madison, WI) in frame with the His6 tag and under the bacteriophage T7 promoter present in the vector. .. Pellets were resuspended in 8 m urea, 0.1 m NaH2 PO4 , 0.01 m Tris, pH 8, 7.5 m m β-mercaptoethanol, and cells were lysed completely by 2 × 30 sec sonication.

    Recombinant:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes. .. Recombinant plasmid p15-16A (aa 17 to 247) was isolated from ampicillin-resistant clones using a ZR plasmid miniprep kit (Zymo Research, Irvine, CA).

    Article Title: Amino Acids Important for DNA Recognition by the Response Regulator OmpR *Amino Acids Important for DNA Recognition by the Response Regulator OmpR * S⃞
    Article Snippet: Overexpression and Purification of OmpR Mutants —To overproduce the mutant OmpR proteins, each plasmid was cleaved with NdeI and BamHI and ligated into the expression vector pET15b (Novagen) that had been cleaved with the same enzymes. .. The ligation mixture was used to transform E. coli strain DH5α, and individual transformants were screened for the presence of the appropriate recombinant plasmid.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM. .. E . coli DH5α was then transformed with the ligation mix and the transformants were selected on Luria agar with ampicillin (100 μg mL-1 ).

    Article Title: Crystallization and preliminary X-ray analysis of a dye-linked d-lactate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix
    Article Snippet: Paragraph title: 2.1. Construction of the expression system and purification of the recombinant protein ... The amplified 1.4 kbp fragment was digested with Nde I and Bam HI and then ligated with the expression vector pET15b (Novagen, Madison, Wisconsin, USA) previously linearized using Nde I and Bam HI, yielding pDLDH.

    DNA Extraction:

    Article Title: Crystallization and preliminary X-ray analysis of a dye-linked d-lactate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix
    Article Snippet: The genomic DNA was prepared using a Genomic DNA Isolation Kit for Bacteria (Nexttec GmbH Biotechnologie, Leverkusen, Germany). .. The amplified 1.4 kbp fragment was digested with Nde I and Bam HI and then ligated with the expression vector pET15b (Novagen, Madison, Wisconsin, USA) previously linearized using Nde I and Bam HI, yielding pDLDH.

    Mutagenesis:

    Article Title: Amino Acids Important for DNA Recognition by the Response Regulator OmpR *Amino Acids Important for DNA Recognition by the Response Regulator OmpR * S⃞
    Article Snippet: .. Overexpression and Purification of OmpR Mutants —To overproduce the mutant OmpR proteins, each plasmid was cleaved with NdeI and BamHI and ligated into the expression vector pET15b (Novagen) that had been cleaved with the same enzymes. .. The ligation mixture was used to transform E. coli strain DH5α, and individual transformants were screened for the presence of the appropriate recombinant plasmid.

    Random Primed:

    Article Title: Characterization of Drosophila Tyramine β-HydroxylaseGene and Isolation of Mutant Flies Lacking Octopamine
    Article Snippet: Subsequently, the filter was hybridized with a random-primed DNA probe of pRP49 ( ) as a control. .. The 1.2 kb Sal – Xho fragment of the DmDBH cDNA (Fig. B ) was inserted into the Xho I site of the bacterial expression vector pET15b (Novagen, Madison, WI) in frame with the His6 tag and under the bacteriophage T7 promoter present in the vector.

    Subcloning:

    Article Title: Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin ▿ Bacteriophage and Its Cloned Biologically Active Endolysin ▿ †
    Article Snippet: Paragraph title: Subcloning of bacteriophage endolysin genes into E. coli and L. lactis . ... NdeI- and XhoI-restricted PCR products were ligated into the expression vector pET15b (Novagen) by using Fast-Link DNA ligase (Epicentre), and the resulting construct, pET15b- cd27l , was introduced into chemically competent E. coli TOP10 (Invitrogen).

    Size-exclusion Chromatography:

    Article Title: Characterization of Drosophila Tyramine β-HydroxylaseGene and Isolation of Mutant Flies Lacking Octopamine
    Article Snippet: The 1.2 kb Sal – Xho fragment of the DmDBH cDNA (Fig. B ) was inserted into the Xho I site of the bacterial expression vector pET15b (Novagen, Madison, WI) in frame with the His6 tag and under the bacteriophage T7 promoter present in the vector. .. Pellets were resuspended in 8 m urea, 0.1 m NaH2 PO4 , 0.01 m Tris, pH 8, 7.5 m m β-mercaptoethanol, and cells were lysed completely by 2 × 30 sec sonication.

    Labeling:

    Article Title: Structural Insight into the Recognition of r(UAG) by Musashi-1 RBD2, and Construction of a Model of Musashi-1 RBD1-2 Bound to the Minimum Target RNA
    Article Snippet: Protein and RNA Preparation The RBD2+, comprising mouse Msi1 residues Lys109-Arg200, containing RBD2, was cloned into the expression vector pET15b (Novagen), as a fusion with an N-terminal hexahistidine affinity tag and thrombin cleavage site. .. The former and latter conditions were used to prepare 15 N-single labeled and 13 C/15 N-doubly labeled proteins, respectively.

    Purification:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: .. The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes. .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Article Title: Amino Acids Important for DNA Recognition by the Response Regulator OmpR *Amino Acids Important for DNA Recognition by the Response Regulator OmpR * S⃞
    Article Snippet: .. Overexpression and Purification of OmpR Mutants —To overproduce the mutant OmpR proteins, each plasmid was cleaved with NdeI and BamHI and ligated into the expression vector pET15b (Novagen) that had been cleaved with the same enzymes. .. The ligation mixture was used to transform E. coli strain DH5α, and individual transformants were screened for the presence of the appropriate recombinant plasmid.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM. .. E . coli DH5α was then transformed with the ligation mix and the transformants were selected on Luria agar with ampicillin (100 μg mL-1 ).

    Article Title: Crystallization and preliminary X-ray analysis of a dye-linked d-lactate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix
    Article Snippet: Paragraph title: 2.1. Construction of the expression system and purification of the recombinant protein ... The amplified 1.4 kbp fragment was digested with Nde I and Bam HI and then ligated with the expression vector pET15b (Novagen, Madison, Wisconsin, USA) previously linearized using Nde I and Bam HI, yielding pDLDH.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. Cloning, overexpression and purification of metagenomic phosphodiesterase A construct for heterologous expression of the gene encoding metagenomic phosphodiesterase (PdeM) was constructed by PCR amplification from the fosmid clone and cloning into the expression vector pET15b (Novagen). .. The complete coding region of the PdeM gene was amplified by PCR using primers pdemF/pdemR designed from the annotated gene sequence (Froward 5’-GGGAATTC CATATG AGCCCCCAAGCTTCACCG-3’ and Reverse 5’-CGC GGATCC TCAAGTTCCCGGCAAGGCCC-3’).

    Protein Purification:

    Article Title: Characterization of Drosophila Tyramine β-HydroxylaseGene and Isolation of Mutant Flies Lacking Octopamine
    Article Snippet: The 1.2 kb Sal – Xho fragment of the DmDBH cDNA (Fig. B ) was inserted into the Xho I site of the bacterial expression vector pET15b (Novagen, Madison, WI) in frame with the His6 tag and under the bacteriophage T7 promoter present in the vector. .. The 1.2 kb Sal – Xho fragment of the DmDBH cDNA (Fig. B ) was inserted into the Xho I site of the bacterial expression vector pET15b (Novagen, Madison, WI) in frame with the His6 tag and under the bacteriophage T7 promoter present in the vector.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Identification of functionally relevant histidine residues in the apoptotic nuclease CAD
    Article Snippet: To isolate the cDNA for human ICAD-L/DFF45 (hICAD-L), total RNA was extracted from HeLa cells with a RNeasy Mini Kit (Qiagen) and used for first-strand cDNA synthesis and subsequent amplification of the hICAD-L gene with the Access RT–PCR system (Promega). .. The coding region for hICAD-L (DFF45) was inserted in-frame as an Nco I– Sal I fragment into the Nco I and Xho I sites of the bacterial expression vector pET15b (Novagen).

    IA:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The cDNA amplification was carried out according to a standard laboratory PCR protocol using forward primer cx16-15F (5′-GCG CTCGAG GCGTTGCTGACTCTGCAAGG-3′) and reverse primer cx16-15R (5′-GC GGATCC GCACTGCCACTGTAACTGAT-3′) (IDT, Coralville, IA). .. The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes.

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin ▿ Bacteriophage and Its Cloned Biologically Active Endolysin ▿ †
    Article Snippet: The ΦCD27 endolysin gene cd27l was amplified from genomic DNA by using primers designed to create an NdeI site at the 5′ end (5′-TTA CAT ATG AAA ATA TGT ATA ACA GTA GG, where underlining indicates the bases corresponding to the restriction endonuclease site) and a XhoI site downstream of the coding sequence (5′-CAA CCA C CT CGA G TT GAT AAC). .. NdeI- and XhoI-restricted PCR products were ligated into the expression vector pET15b (Novagen) by using Fast-Link DNA ligase (Epicentre), and the resulting construct, pET15b- cd27l , was introduced into chemically competent E. coli TOP10 (Invitrogen).

    Plasmid Preparation:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: .. The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes. .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Article Title: Sublingual Immunization with M2-Based Vaccine Induces Broad Protective Immunity against Influenza
    Article Snippet: .. For the plasmid expressing 3M2eC, the gene was digested with Xho I and BamH I and inserted into the bacterial expression vector pET15b (Novagen, Madison, WI) to express as a fusion of his-tag at the N terminus, resulting in the plasmid pET15b-3M2eC. .. For the construct expressing M2eC protein, which has one M2e domain, the synthesized gene was digested with Nde I and BamH I, and then inserted into pET15b vector.

    Article Title: Structural Insight into the Recognition of r(UAG) by Musashi-1 RBD2, and Construction of a Model of Musashi-1 RBD1-2 Bound to the Minimum Target RNA
    Article Snippet: .. Protein and RNA Preparation The RBD2+, comprising mouse Msi1 residues Lys109-Arg200, containing RBD2, was cloned into the expression vector pET15b (Novagen), as a fusion with an N-terminal hexahistidine affinity tag and thrombin cleavage site. ..

    Article Title: Amino Acids Important for DNA Recognition by the Response Regulator OmpR *Amino Acids Important for DNA Recognition by the Response Regulator OmpR * S⃞
    Article Snippet: .. Overexpression and Purification of OmpR Mutants —To overproduce the mutant OmpR proteins, each plasmid was cleaved with NdeI and BamHI and ligated into the expression vector pET15b (Novagen) that had been cleaved with the same enzymes. .. The ligation mixture was used to transform E. coli strain DH5α, and individual transformants were screened for the presence of the appropriate recombinant plasmid.

    Article Title: Molecular Characterization of a Novel Glucosyltransferase from Lactobacillus reuteri Strain 121 Synthesizing a Unique, Highly Branched Glucan with ?-(1- > 4) and ?-(1- > 6) Glucosidic Bonds
    Article Snippet: .. Plasmid pBGTF1 was digested with Nco I/ Bam HI, and the resulting 5.3-kb fragment was ligated into the corresponding sites of the expression vector pET15b (Novagen) yielding p15gtf. ..

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM. .. E . coli DH5α was then transformed with the ligation mix and the transformants were selected on Luria agar with ampicillin (100 μg mL-1 ).

    Article Title: Crystallization and preliminary X-ray analysis of a dye-linked d-lactate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix
    Article Snippet: .. The amplified 1.4 kbp fragment was digested with Nde I and Bam HI and then ligated with the expression vector pET15b (Novagen, Madison, Wisconsin, USA) previously linearized using Nde I and Bam HI, yielding pDLDH. .. E. coli strain Rosetta-gami 2 (DE3) (Stratagene, La Jolla, California, USA) was then transformed with the vector and the transformants were cultivated at 310 K in 1 l SB medium (1.2% tryptone peptone, 2.4% yeast extract, 1.25% K2 HPO4 , 0.38% KH2 PO4 and 0.5% glycerol) containing 50 µg ml−1 ampicillin and 50 µg ml−1 chloramphenicol until the optical density at 600 nm reached 0.6.

    Article Title: Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin ▿ Bacteriophage and Its Cloned Biologically Active Endolysin ▿ †
    Article Snippet: .. NdeI- and XhoI-restricted PCR products were ligated into the expression vector pET15b (Novagen) by using Fast-Link DNA ligase (Epicentre), and the resulting construct, pET15b- cd27l , was introduced into chemically competent E. coli TOP10 (Invitrogen). .. Positive transformants were selected with ampicillin (100 μg/ml), gene identity was confirmed by sequencing, and chemically competent E. coli BL21(DE3) (Invitrogen) was transformed with the plasmid constructs or the original vector pET15b for protein expression.

    Article Title: Identification of functionally relevant histidine residues in the apoptotic nuclease CAD
    Article Snippet: .. The coding region for hICAD-L (DFF45) was inserted in-frame as an Nco I– Sal I fragment into the Nco I and Xho I sites of the bacterial expression vector pET15b (Novagen). ..

    Article Title: Characterization of Drosophila Tyramine β-HydroxylaseGene and Isolation of Mutant Flies Lacking Octopamine
    Article Snippet: .. The 1.2 kb Sal – Xho fragment of the DmDBH cDNA (Fig. B ) was inserted into the Xho I site of the bacterial expression vector pET15b (Novagen, Madison, WI) in frame with the His6 tag and under the bacteriophage T7 promoter present in the vector. .. The resulting plasmid was introduced into the Escherichia coli BL21(DE3) strain ( ); cultures of the transformed cells were grown to OD600 = 0.5, induced with 1 m m isopropyl β- d -thiogalactopyranoside, then grown for 3 more hr and harvested.

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. Cloning, overexpression and purification of metagenomic phosphodiesterase A construct for heterologous expression of the gene encoding metagenomic phosphodiesterase (PdeM) was constructed by PCR amplification from the fosmid clone and cloning into the expression vector pET15b (Novagen). .. The complete coding region of the PdeM gene was amplified by PCR using primers pdemF/pdemR designed from the annotated gene sequence (Froward 5’-GGGAATTC CATATG AGCCCCCAAGCTTCACCG-3’ and Reverse 5’-CGC GGATCC TCAAGTTCCCGGCAAGGCCC-3’).

    Positron Emission Tomography:

    Article Title: Identification and Characterization of a Novel Phosphodiesterase from the Metagenome of an Indian Coalbed
    Article Snippet: .. The amplicon of 747 bp was digested with NdeI/BamHI restriction enzymes, purified with PCR purification kit (Promega) and finally ligated with the similarly digested expression vector pET15b (Novagen) to generate the recombinant plasmid, pET-PdeM. .. E . coli DH5α was then transformed with the ligation mix and the transformants were selected on Luria agar with ampicillin (100 μg mL-1 ).

    Agarose Gel Electrophoresis:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The amplified EqCXCL16 fragment (amino acids [aa] 17 to 247) was run on a 1% agarose gel (E-Gel EX; Life Technologies, Grand Island, NY) and purified with a commercial kit (Zymoclean gel recovery kit; Zymo Research, Irvine, CA). .. The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes.

    In Situ:

    Article Title: Characterization of Drosophila Tyramine β-HydroxylaseGene and Isolation of Mutant Flies Lacking Octopamine
    Article Snippet: In situ detection of RNA in whole-mount larval CNS was as described in using digoxygenin-labeled DNA probe synthesized for the Pst – Xho fragment of the cDNA clone (Fig. B ) according to the manufacturer’s protocols (Boehringer Mannheim, Indianapolis, IN). .. The 1.2 kb Sal – Xho fragment of the DmDBH cDNA (Fig. B ) was inserted into the Xho I site of the bacterial expression vector pET15b (Novagen, Madison, WI) in frame with the His6 tag and under the bacteriophage T7 promoter present in the vector.

    Produced:

    Article Title: Identification of functionally relevant histidine residues in the apoptotic nuclease CAD
    Article Snippet: The open reading frame for mCAD was inserted in-frame into the Nco I and Not I sites of pTriEx1 (Novagen) such that a coding region for an N-terminal His6 tag was produced at the 5′-end of the mCAD gene, resulting in plasmid pHis-mCAD. .. The coding region for hICAD-L (DFF45) was inserted in-frame as an Nco I– Sal I fragment into the Nco I and Xho I sites of the bacterial expression vector pET15b (Novagen).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92
    Millipore expression vector pet15b
    Expression Vector Pet15b, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 113 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/expression vector pet15b/product/Millipore
    Average 92 stars, based on 113 article reviews
    Price from $9.99 to $1999.99
    expression vector pet15b - by Bioz Stars, 2020-04
    92/100 stars
      Buy from Supplier

    86
    Millipore t7 expression plasmids pet15b
    T7 Expression Plasmids Pet15b, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 expression plasmids pet15b/product/Millipore
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t7 expression plasmids pet15b - by Bioz Stars, 2020-04
    86/100 stars
      Buy from Supplier

    87
    Millipore pet15b protein expression vector
    Pet15b Protein Expression Vector, supplied by Millipore, used in various techniques. Bioz Stars score: 87/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pet15b protein expression vector/product/Millipore
    Average 87 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pet15b protein expression vector - by Bioz Stars, 2020-04
    87/100 stars
      Buy from Supplier

    Image Search Results