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Promega puc19
Autoradiograph of an SDS–12% polyacrylamide gel containing [ 35 S]methionine-labelled products of in vitro transcription-translation. Lanes: 1, pGEM-T Easy; 2, pTK3251, containing the A. actinomycetemcomitans cdtA gene; 3, pTK3252, containing the A. actinomycetemcomitans cdtB gene; 4, pTK3253, containing the A. actinomycetemcomitans cdtC gene; 5, pTK3022, containing the entire cdtABC gene cluster; 6, <t>pUC19.</t> The CDT activities of sterile sonic lysates from the recombinant strains are indicated at the bottom. Radiolabelled bands marked by #, ∗, and + are putative gene products of cdtA , - B , and - C , respectively.
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1) Product Images from "The Cell Cycle-Specific Growth-Inhibitory Factor Produced by Actinobacillus actinomycetemcomitans Is a Cytolethal Distending Toxin"

Article Title: The Cell Cycle-Specific Growth-Inhibitory Factor Produced by Actinobacillus actinomycetemcomitans Is a Cytolethal Distending Toxin

Journal: Infection and Immunity

doi:

Autoradiograph of an SDS–12% polyacrylamide gel containing [ 35 S]methionine-labelled products of in vitro transcription-translation. Lanes: 1, pGEM-T Easy; 2, pTK3251, containing the A. actinomycetemcomitans cdtA gene; 3, pTK3252, containing the A. actinomycetemcomitans cdtB gene; 4, pTK3253, containing the A. actinomycetemcomitans cdtC gene; 5, pTK3022, containing the entire cdtABC gene cluster; 6, pUC19. The CDT activities of sterile sonic lysates from the recombinant strains are indicated at the bottom. Radiolabelled bands marked by #, ∗, and + are putative gene products of cdtA , - B , and - C , respectively.
Figure Legend Snippet: Autoradiograph of an SDS–12% polyacrylamide gel containing [ 35 S]methionine-labelled products of in vitro transcription-translation. Lanes: 1, pGEM-T Easy; 2, pTK3251, containing the A. actinomycetemcomitans cdtA gene; 3, pTK3252, containing the A. actinomycetemcomitans cdtB gene; 4, pTK3253, containing the A. actinomycetemcomitans cdtC gene; 5, pTK3022, containing the entire cdtABC gene cluster; 6, pUC19. The CDT activities of sterile sonic lysates from the recombinant strains are indicated at the bottom. Radiolabelled bands marked by #, ∗, and + are putative gene products of cdtA , - B , and - C , respectively.

Techniques Used: Autoradiography, In Vitro, Recombinant

Related Articles

Clone Assay:

Article Title: Intracellular trafficking of nucleic acids
Article Snippet: In these non-dividing cells, whereas SV40 DNA localised in the nuclei with 8 h, other plasmids, including pBR322, pUC19 and pGL3-basic (Promega Corp.), did not enter the nuclei until the cells were allowed to divide [ , ]. .. Indeed, when as little as 50 bp of the SV40 enhancer was cloned into these other plasmids, they were able to enter nuclei with the same kinetics as the entire SV40 genome [ , ].

Article Title: The cysteine desulfurase, IscS, has a major role in in vivo Fe-S cluster formation in Escherichia coli
Article Snippet: .. Plasmids pPK4071 and pPK4083 were made by digesting λ430 ( ) with Nru I and cloning this 5-kb fragment containing iscRSUAhscB ( ) into the Sma I site of pUC19 ( ) and pAlter-1 (Promega), respectively. .. Plasmids pPK4196 and pPK4194 were generated by cloning PCR products containing only the ORFs for iscS and iscSUA into pET11a , respectively (Fig. ).

Article Title: Insights into functional and evolutionary analysis of carbaryl metabolic pathway from Pseudomonas sp. strain C5pp
Article Snippet: .. One of the randomly chosen clone designated as G1 was digested with Bam HI and all six fragments were sub-cloned into pUC19 and partially sequenced using universal M13 forward and reverse primers (M13F, 5′-GTTTTCCCAGTCACGAC-3′ and M13R, 5′-CAGGAAACAGCTATGAC-3′, Promega, USA) after cloning it into pUC19 at Bam HI site. ..

Article Title: Cloning, Characterization, Controlled Overexpression, and Inactivation of the Major Tributyrin Esterase Gene of Lactococcus lactis
Article Snippet: .. The plasmid vectors used in the cloning experiments with E. coli were pUC18 and pUC19 ( ) and pGEM-5Zf(+) (Promega Corp.). ..

Article Title: Measurement of Biologically Available Naphthalene in Gas and Aqueous Phases by Use of a Pseudomonas putida Biosensor
Article Snippet: .. The plasmids used during this study included pUC19 , for general subclonings; pGEM-T-Easy (Promega Corp.), for cloning of DNA fragments generated in PCRs; pUC18Not , for providing Not I symmetrical restriction sites; and PCK218 , serving as the mini-Tn 5 vehicle to deliver the nahR-nahG ′:: luxAB construct to the chromosome of P. putida pPG7. .. A 1.1-kb fragment containing the nahR gene and the sal promoter was recovered from plasmid pCNB4- lacZ ( ) by digestion with Pst I and treatment with T4 DNA polymerase and, subsequently, digestion with Eco RI and treatment with Klenow DNA polymerase.

Article Title: Host Ranges of Listeria-Specific Bacteriophages from the Turkey Processing Plant Environment in the United States ▿
Article Snippet: .. The DNA was digested with NheI (New England Biolabs, Waverly, MA) and cloned into pUC19 (Promega, Madison, WI) digested with XbaI (New England Biolabs). .. The recombinant plasmids were transformed into competent Escherichia coli DH5α cells, and transformants were selected using ampicillin (100 μg/ml) and X-Gal (5-bromo-4-chloro-3-indolyl-β- d ).

Article Title: Tn5 Transposase with an Altered Specificity for Transposon Ends
Article Snippet: The full-length Eco RI- Kpn I transposon fragment was ligated into pUC19 to create pRZ1495/pUC. pRZ1495/pUC was digested with Not I and Afl II, filled in with dNTPs and T4 DNA polymerase (Promega), and religated to form pRZΔ1495/pUC. .. The Eco RI/ Kpn I transposon-containing fragment from pRZΔ1495/pUC was then cloned into the multicloning site of pAltEX2 (Promega) to form pRZ9904 (OE/OE).

Article Title: Intracellular Glycerol Levels Modulate the Activity of Sln1p, a Saccharomyces cerevisiae Two-component Regulator *
Article Snippet: .. A 1.9-kb Ava I- Bam HI restriction fragment was cloned into pUC19 (Promega Biotech) to generate the intermediate plasmid, pJF1068. .. A deletion of 268 base pairs was introduced into the GPD1 open reading frame on pJF1068 by Sal I digestion and ligation.

Amplification:

Article Title: The cysteine desulfurase, IscS, has a major role in in vivo Fe-S cluster formation in Escherichia coli
Article Snippet: Plasmids pPK4071 and pPK4083 were made by digesting λ430 ( ) with Nru I and cloning this 5-kb fragment containing iscRSUAhscB ( ) into the Sma I site of pUC19 ( ) and pAlter-1 (Promega), respectively. .. For DNA amplification by PCR from chromosomal or plasmid DNA, either Pfu DNA polymerase (Stratagene) or Taq DNA polymerase (Promega) was used.

Article Title: Quantifying promoter activity during the developmental cycle of Chlamydia trachomatis
Article Snippet: The 5′ end of the gfp transcript, which is initiated by P3, was then amplified with random-primer reverse transcription and nested PCR using gfp specific primers with the provided primers in the kit. .. PCR products were then inserted into pUC19 (Promega, Madison, WI) for DNA sequencing.

Article Title: Cap-Independent Translational Enhancement of Turnip Crinkle Virus Genomic and Subgenomic RNAs
Article Snippet: .. The sgUTR-luc-3′TCV construct was made as follows: (i) the cDNA of the 1.45-kb sgRNA was PCR amplified from mutant mprNco by using one oligodeoxyribonucleotide containing the T7 promoter sequence as its 5′ half and TCV sequence from nucleotide (nt) 2607 to 2630 as its 3′ half and another oligodeoxyribonucleotide with sequence complementary to TCV nt 4054 to 4030; (ii) the PCR-amplified fragment was cloned into pUC19, and a Sna BI site was introduced at the 3′ end of the TCV CP coding region; (iii) the resulting plasmid was cut with Nco I and Sna BI, and the CP coding region was replaced with the firefly luciferase gene ( luc ) obtained by cutting pSP-luc+ (Promega, Madison, Wis.) with Nco I and Xba I. .. All other luc -containing constructs were derived from the sgUTR-luc-3′TCV by manipulating either the 5′ or 3′ UTR region or both.

Article Title: Intracellular Glycerol Levels Modulate the Activity of Sln1p, a Saccharomyces cerevisiae Two-component Regulator *
Article Snippet: A GPD1 fragment encompassing the entire open reading frame was polymerase chain reaction amplified using a primer pair corresponding to open reading frame YDL022W (Research Genetics) and genomic DNA as template. .. A 1.9-kb Ava I- Bam HI restriction fragment was cloned into pUC19 (Promega Biotech) to generate the intermediate plasmid, pJF1068.

Construct:

Article Title: Insights into functional and evolutionary analysis of carbaryl metabolic pathway from Pseudomonas sp. strain C5pp
Article Snippet: The genomic DNA library of Pseudomonas sp. strain C5pp was constructed in CopyControl fosmid pCC2FOS with a phage titer of 3 × 106 CFU.ml−1 and a pool of ~9600 colonies was used for screening. .. One of the randomly chosen clone designated as G1 was digested with Bam HI and all six fragments were sub-cloned into pUC19 and partially sequenced using universal M13 forward and reverse primers (M13F, 5′-GTTTTCCCAGTCACGAC-3′ and M13R, 5′-CAGGAAACAGCTATGAC-3′, Promega, USA) after cloning it into pUC19 at Bam HI site.

Article Title: Cap-Independent Translational Enhancement of Turnip Crinkle Virus Genomic and Subgenomic RNAs
Article Snippet: .. The sgUTR-luc-3′TCV construct was made as follows: (i) the cDNA of the 1.45-kb sgRNA was PCR amplified from mutant mprNco by using one oligodeoxyribonucleotide containing the T7 promoter sequence as its 5′ half and TCV sequence from nucleotide (nt) 2607 to 2630 as its 3′ half and another oligodeoxyribonucleotide with sequence complementary to TCV nt 4054 to 4030; (ii) the PCR-amplified fragment was cloned into pUC19, and a Sna BI site was introduced at the 3′ end of the TCV CP coding region; (iii) the resulting plasmid was cut with Nco I and Sna BI, and the CP coding region was replaced with the firefly luciferase gene ( luc ) obtained by cutting pSP-luc+ (Promega, Madison, Wis.) with Nco I and Xba I. .. All other luc -containing constructs were derived from the sgUTR-luc-3′TCV by manipulating either the 5′ or 3′ UTR region or both.

Article Title: Inhibition of Hepatitis B Virus Replication by MyD88 Involves Accelerated Degradation of Pregenomic RNA and Nuclear Retention of Pre-S/S RNAs ▿
Article Snippet: .. To construct HBV plasmid pHBV1.3, a terminally redundant (1.3× copy), replication-competent HBV genome (subtype adw; nucleotides 957 to 1952; GenBank accession number ) was inserted into pUC19 (Promega). pCIdA-HBV expresses all the HBV proteins, but it lacks the 5′ ɛ RNA signal, which eliminates pregenomic RNA packaging and DNA replication ( ). .. Both pCMV-HBV ( ) and pTet-HBV ( ) contain the wild-type HBV 1.1-mer overlength genomic sequence.

Article Title: Measurement of Biologically Available Naphthalene in Gas and Aqueous Phases by Use of a Pseudomonas putida Biosensor
Article Snippet: .. The plasmids used during this study included pUC19 , for general subclonings; pGEM-T-Easy (Promega Corp.), for cloning of DNA fragments generated in PCRs; pUC18Not , for providing Not I symmetrical restriction sites; and PCK218 , serving as the mini-Tn 5 vehicle to deliver the nahR-nahG ′:: luxAB construct to the chromosome of P. putida pPG7. .. A 1.1-kb fragment containing the nahR gene and the sal promoter was recovered from plasmid pCNB4- lacZ ( ) by digestion with Pst I and treatment with T4 DNA polymerase and, subsequently, digestion with Eco RI and treatment with Klenow DNA polymerase.

Article Title: Tn5 Transposase with an Altered Specificity for Transposon Ends
Article Snippet: The papillation vector, pRZ9904 (IE12A/IE12A), was constructed in the following manner. pRZ1495 ( ) was digested to completion with Kpn I and partially digested with Eco RI. .. The full-length Eco RI- Kpn I transposon fragment was ligated into pUC19 to create pRZ1495/pUC. pRZ1495/pUC was digested with Not I and Afl II, filled in with dNTPs and T4 DNA polymerase (Promega), and religated to form pRZΔ1495/pUC.

Luciferase:

Article Title: Cap-Independent Translational Enhancement of Turnip Crinkle Virus Genomic and Subgenomic RNAs
Article Snippet: .. The sgUTR-luc-3′TCV construct was made as follows: (i) the cDNA of the 1.45-kb sgRNA was PCR amplified from mutant mprNco by using one oligodeoxyribonucleotide containing the T7 promoter sequence as its 5′ half and TCV sequence from nucleotide (nt) 2607 to 2630 as its 3′ half and another oligodeoxyribonucleotide with sequence complementary to TCV nt 4054 to 4030; (ii) the PCR-amplified fragment was cloned into pUC19, and a Sna BI site was introduced at the 3′ end of the TCV CP coding region; (iii) the resulting plasmid was cut with Nco I and Sna BI, and the CP coding region was replaced with the firefly luciferase gene ( luc ) obtained by cutting pSP-luc+ (Promega, Madison, Wis.) with Nco I and Xba I. .. All other luc -containing constructs were derived from the sgUTR-luc-3′TCV by manipulating either the 5′ or 3′ UTR region or both.

Article Title: Inhibition of Hepatitis B Virus Replication by MyD88 Involves Accelerated Degradation of Pregenomic RNA and Nuclear Retention of Pre-S/S RNAs ▿
Article Snippet: To construct HBV plasmid pHBV1.3, a terminally redundant (1.3× copy), replication-competent HBV genome (subtype adw; nucleotides 957 to 1952; GenBank accession number ) was inserted into pUC19 (Promega). pCIdA-HBV expresses all the HBV proteins, but it lacks the 5′ ɛ RNA signal, which eliminates pregenomic RNA packaging and DNA replication ( ). .. The luciferase reporter constructs of HBV promoters/enhancers (ENII/Cp, Sp1, Sp2, and ENI/Xp) were described previously ( ).

Activity Assay:

Article Title: Intracellular trafficking of nucleic acids
Article Snippet: In these non-dividing cells, whereas SV40 DNA localised in the nuclei with 8 h, other plasmids, including pBR322, pUC19 and pGL3-basic (Promega Corp.), did not enter the nuclei until the cells were allowed to divide [ , ]. .. This finding supports earlier studies by Graessman [ ] in which the SV40 enhancer was postulated to have a ‘helper’ activity for nuclear localisation apart from the classical transcriptional activity attributed to it.

Modification:

Article Title: Intracellular Glycerol Levels Modulate the Activity of Sln1p, a Saccharomyces cerevisiae Two-component Regulator *
Article Snippet: The modified MCM1 gene was then subcloned as a 3.5-kb Xba I- Xho I fragment into pRS316 ( URA3 , CEN ) ( ). .. A 1.9-kb Ava I- Bam HI restriction fragment was cloned into pUC19 (Promega Biotech) to generate the intermediate plasmid, pJF1068.

Transformation Assay:

Article Title: Host Ranges of Listeria-Specific Bacteriophages from the Turkey Processing Plant Environment in the United States ▿
Article Snippet: The DNA was digested with NheI (New England Biolabs, Waverly, MA) and cloned into pUC19 (Promega, Madison, WI) digested with XbaI (New England Biolabs). .. The recombinant plasmids were transformed into competent Escherichia coli DH5α cells, and transformants were selected using ampicillin (100 μg/ml) and X-Gal (5-bromo-4-chloro-3-indolyl-β- d ).

Over Expression:

Article Title: Molecular Cloning, Expression of minD Gene from Lactobacillus acidophilus VTCC-B-871 and Analyses to Identify Lactobacillus rhamnosus PN04 from Vietnam Hottuynia cordata Thunb.
Article Snippet: The pUC19 and pGEM-T vectors used for molecular cloning and E. coli JM109, BL21(DE3)pLysS were purchased by Promega. .. The pET28 (a+) used for overexpression was purchased by Novagen.

Derivative Assay:

Article Title: Cap-Independent Translational Enhancement of Turnip Crinkle Virus Genomic and Subgenomic RNAs
Article Snippet: The sgUTR-luc-3′TCV construct was made as follows: (i) the cDNA of the 1.45-kb sgRNA was PCR amplified from mutant mprNco by using one oligodeoxyribonucleotide containing the T7 promoter sequence as its 5′ half and TCV sequence from nucleotide (nt) 2607 to 2630 as its 3′ half and another oligodeoxyribonucleotide with sequence complementary to TCV nt 4054 to 4030; (ii) the PCR-amplified fragment was cloned into pUC19, and a Sna BI site was introduced at the 3′ end of the TCV CP coding region; (iii) the resulting plasmid was cut with Nco I and Sna BI, and the CP coding region was replaced with the firefly luciferase gene ( luc ) obtained by cutting pSP-luc+ (Promega, Madison, Wis.) with Nco I and Xba I. .. All other luc -containing constructs were derived from the sgUTR-luc-3′TCV by manipulating either the 5′ or 3′ UTR region or both.

Hybridization:

Article Title: Intracellular trafficking of nucleic acids
Article Snippet: When protein-free, purified SV40 DNA (5243 bp) was microinjected into the cytoplasm of TC7 African Green monkey kidney cells, the majority of plasmids were detected in cell nuclei by 6 – 8 h after injection by in situ hybridisation. .. In these non-dividing cells, whereas SV40 DNA localised in the nuclei with 8 h, other plasmids, including pBR322, pUC19 and pGL3-basic (Promega Corp.), did not enter the nuclei until the cells were allowed to divide [ , ].

Conjugation Assay:

Article Title: Measurement of Biologically Available Naphthalene in Gas and Aqueous Phases by Use of a Pseudomonas putida Biosensor
Article Snippet: E. coli CC118λpir was used for the propagation of plasmids with an R6K origin of replication, and E. coli HB101(pRK2013) was used as a helper strain, providing transfer functions during conjugation. .. The plasmids used during this study included pUC19 , for general subclonings; pGEM-T-Easy (Promega Corp.), for cloning of DNA fragments generated in PCRs; pUC18Not , for providing Not I symmetrical restriction sites; and PCK218 , serving as the mini-Tn 5 vehicle to deliver the nahR-nahG ′:: luxAB construct to the chromosome of P. putida pPG7.

Ligation:

Article Title: Intracellular Glycerol Levels Modulate the Activity of Sln1p, a Saccharomyces cerevisiae Two-component Regulator *
Article Snippet: A 1.9-kb Ava I- Bam HI restriction fragment was cloned into pUC19 (Promega Biotech) to generate the intermediate plasmid, pJF1068. .. A deletion of 268 base pairs was introduced into the GPD1 open reading frame on pJF1068 by Sal I digestion and ligation.

Cell Culture:

Article Title: The Cell Cycle-Specific Growth-Inhibitory Factor Produced by Actinobacillus actinomycetemcomitans Is a Cytolethal Distending Toxin
Article Snippet: A. actinomycetemcomitans Y4 (serotype b, ATCC 43718) was cultured in Trypticase soy broth (Becton Dickinson Microbiology Systems, Cockeysville, Md.) supplemented with 1% (wt/vol) yeast extract in a 5% CO2 atmosphere. .. Manipulation of DNA in E. coli was carried out with pUC19 , pGEM-T Easy (Promega, Madison, Wis.), or pET-28a(+) (Novagen, Madison, Wis.).

Generated:

Article Title: The cysteine desulfurase, IscS, has a major role in in vivo Fe-S cluster formation in Escherichia coli
Article Snippet: Plasmids pPK4071 and pPK4083 were made by digesting λ430 ( ) with Nru I and cloning this 5-kb fragment containing iscRSUAhscB ( ) into the Sma I site of pUC19 ( ) and pAlter-1 (Promega), respectively. .. Plasmids pPK4196 and pPK4194 were generated by cloning PCR products containing only the ORFs for iscS and iscSUA into pET11a , respectively (Fig. ).

Article Title: Measurement of Biologically Available Naphthalene in Gas and Aqueous Phases by Use of a Pseudomonas putida Biosensor
Article Snippet: .. The plasmids used during this study included pUC19 , for general subclonings; pGEM-T-Easy (Promega Corp.), for cloning of DNA fragments generated in PCRs; pUC18Not , for providing Not I symmetrical restriction sites; and PCK218 , serving as the mini-Tn 5 vehicle to deliver the nahR-nahG ′:: luxAB construct to the chromosome of P. putida pPG7. .. A 1.1-kb fragment containing the nahR gene and the sal promoter was recovered from plasmid pCNB4- lacZ ( ) by digestion with Pst I and treatment with T4 DNA polymerase and, subsequently, digestion with Eco RI and treatment with Klenow DNA polymerase.

DNA Sequencing:

Article Title: Quantifying promoter activity during the developmental cycle of Chlamydia trachomatis
Article Snippet: .. PCR products were then inserted into pUC19 (Promega, Madison, WI) for DNA sequencing. .. Real-time reverse transcription quantitative PCR (RT-qPCR) C. trachomatis infected HeLa cells were harvested at indicated times post infection.

Polymerase Chain Reaction:

Article Title: The cysteine desulfurase, IscS, has a major role in in vivo Fe-S cluster formation in Escherichia coli
Article Snippet: Plasmids pPK4071 and pPK4083 were made by digesting λ430 ( ) with Nru I and cloning this 5-kb fragment containing iscRSUAhscB ( ) into the Sma I site of pUC19 ( ) and pAlter-1 (Promega), respectively. .. Plasmids pPK4196 and pPK4194 were generated by cloning PCR products containing only the ORFs for iscS and iscSUA into pET11a , respectively (Fig. ).

Article Title: Quantifying promoter activity during the developmental cycle of Chlamydia trachomatis
Article Snippet: .. PCR products were then inserted into pUC19 (Promega, Madison, WI) for DNA sequencing. .. Real-time reverse transcription quantitative PCR (RT-qPCR) C. trachomatis infected HeLa cells were harvested at indicated times post infection.

Article Title: Cap-Independent Translational Enhancement of Turnip Crinkle Virus Genomic and Subgenomic RNAs
Article Snippet: .. The sgUTR-luc-3′TCV construct was made as follows: (i) the cDNA of the 1.45-kb sgRNA was PCR amplified from mutant mprNco by using one oligodeoxyribonucleotide containing the T7 promoter sequence as its 5′ half and TCV sequence from nucleotide (nt) 2607 to 2630 as its 3′ half and another oligodeoxyribonucleotide with sequence complementary to TCV nt 4054 to 4030; (ii) the PCR-amplified fragment was cloned into pUC19, and a Sna BI site was introduced at the 3′ end of the TCV CP coding region; (iii) the resulting plasmid was cut with Nco I and Sna BI, and the CP coding region was replaced with the firefly luciferase gene ( luc ) obtained by cutting pSP-luc+ (Promega, Madison, Wis.) with Nco I and Xba I. .. All other luc -containing constructs were derived from the sgUTR-luc-3′TCV by manipulating either the 5′ or 3′ UTR region or both.

Article Title: Intracellular Glycerol Levels Modulate the Activity of Sln1p, a Saccharomyces cerevisiae Two-component Regulator *
Article Snippet: A GPD1 fragment encompassing the entire open reading frame was polymerase chain reaction amplified using a primer pair corresponding to open reading frame YDL022W (Research Genetics) and genomic DNA as template. .. A 1.9-kb Ava I- Bam HI restriction fragment was cloned into pUC19 (Promega Biotech) to generate the intermediate plasmid, pJF1068.

Injection:

Article Title: Intracellular trafficking of nucleic acids
Article Snippet: When protein-free, purified SV40 DNA (5243 bp) was microinjected into the cytoplasm of TC7 African Green monkey kidney cells, the majority of plasmids were detected in cell nuclei by 6 – 8 h after injection by in situ hybridisation. .. In these non-dividing cells, whereas SV40 DNA localised in the nuclei with 8 h, other plasmids, including pBR322, pUC19 and pGL3-basic (Promega Corp.), did not enter the nuclei until the cells were allowed to divide [ , ].

Recombinant:

Article Title: Host Ranges of Listeria-Specific Bacteriophages from the Turkey Processing Plant Environment in the United States ▿
Article Snippet: The DNA was digested with NheI (New England Biolabs, Waverly, MA) and cloned into pUC19 (Promega, Madison, WI) digested with XbaI (New England Biolabs). .. The recombinant plasmids were transformed into competent Escherichia coli DH5α cells, and transformants were selected using ampicillin (100 μg/ml) and X-Gal (5-bromo-4-chloro-3-indolyl-β- d ).

Molecular Cloning:

Article Title: Molecular Cloning, Expression of minD Gene from Lactobacillus acidophilus VTCC-B-871 and Analyses to Identify Lactobacillus rhamnosus PN04 from Vietnam Hottuynia cordata Thunb.
Article Snippet: .. The pUC19 and pGEM-T vectors used for molecular cloning and E. coli JM109, BL21(DE3)pLysS were purchased by Promega. .. The pET28 (a+) used for overexpression was purchased by Novagen.

Mutagenesis:

Article Title: Cap-Independent Translational Enhancement of Turnip Crinkle Virus Genomic and Subgenomic RNAs
Article Snippet: .. The sgUTR-luc-3′TCV construct was made as follows: (i) the cDNA of the 1.45-kb sgRNA was PCR amplified from mutant mprNco by using one oligodeoxyribonucleotide containing the T7 promoter sequence as its 5′ half and TCV sequence from nucleotide (nt) 2607 to 2630 as its 3′ half and another oligodeoxyribonucleotide with sequence complementary to TCV nt 4054 to 4030; (ii) the PCR-amplified fragment was cloned into pUC19, and a Sna BI site was introduced at the 3′ end of the TCV CP coding region; (iii) the resulting plasmid was cut with Nco I and Sna BI, and the CP coding region was replaced with the firefly luciferase gene ( luc ) obtained by cutting pSP-luc+ (Promega, Madison, Wis.) with Nco I and Xba I. .. All other luc -containing constructs were derived from the sgUTR-luc-3′TCV by manipulating either the 5′ or 3′ UTR region or both.

Article Title: Inhibition of Hepatitis B Virus Replication by MyD88 Involves Accelerated Degradation of Pregenomic RNA and Nuclear Retention of Pre-S/S RNAs ▿
Article Snippet: To construct HBV plasmid pHBV1.3, a terminally redundant (1.3× copy), replication-competent HBV genome (subtype adw; nucleotides 957 to 1952; GenBank accession number ) was inserted into pUC19 (Promega). pCIdA-HBV expresses all the HBV proteins, but it lacks the 5′ ɛ RNA signal, which eliminates pregenomic RNA packaging and DNA replication ( ). .. For these plasmids, the synthesis of the pregenomic RNA is driven by the cytomegalovirus (CMV) promoter and the Tet promoter, respectively. pCMV-HBV M2 is a derivative of pCMV-HBV in which the La protein-binding sites have been mutated ( ). pCMV-HBVΔPRE is a PRE deletion mutant of pCMV-HBV ( ).

Article Title: Intracellular Glycerol Levels Modulate the Activity of Sln1p, a Saccharomyces cerevisiae Two-component Regulator *
Article Snippet: The plasmid complemented the α -halo defect of an mcm1-1 mutant. pWT1037 consists of a 2.6-kb Bam HI- Hin dIII fragment containing FPS1 cloned into the Bam HI and Hin dIII sites of the LEU2 CEN vector, pRS315 ( ). pWT1039 consists of a 2.8-kb Hin dIII- Eco RI fragment containing SDH2 and YLL042C cloned into the Hin dIII and Eco RI sites of pRS315. pWT1040 is a derivative of pWT1037 from which an internal 0.9-kb Xho I- Pst I fragment was deleted and replaced with a 3.3-kb Xho I- Pst I LEU2 fragment excised from plasmid YEp13. pJF1070 is a LEU2 marked disruption of the GPD1 gene. .. A 1.9-kb Ava I- Bam HI restriction fragment was cloned into pUC19 (Promega Biotech) to generate the intermediate plasmid, pJF1068.

Isolation:

Article Title: Insights into functional and evolutionary analysis of carbaryl metabolic pathway from Pseudomonas sp. strain C5pp
Article Snippet: Fosmid DNA was isolated by Fosmid Max DNA purification kit (FMAX 046, USA) and restriction digested with Bam HI (~12, 8, 5, 2.8, 2.5 and 1.8 kb) or Not I (20, 11, 5, 4.8, 2.7 and 2.3 kb). .. One of the randomly chosen clone designated as G1 was digested with Bam HI and all six fragments were sub-cloned into pUC19 and partially sequenced using universal M13 forward and reverse primers (M13F, 5′-GTTTTCCCAGTCACGAC-3′ and M13R, 5′-CAGGAAACAGCTATGAC-3′, Promega, USA) after cloning it into pUC19 at Bam HI site.

Article Title: Host Ranges of Listeria-Specific Bacteriophages from the Turkey Processing Plant Environment in the United States ▿
Article Snippet: Phage 20422-1 DNA was purified from phage isolated following infections of L. monocytogenes DP-L862 using the Lambda miniprep phage extraction kit (Qiagen, Valencia, CA). .. The DNA was digested with NheI (New England Biolabs, Waverly, MA) and cloned into pUC19 (Promega, Madison, WI) digested with XbaI (New England Biolabs).

Article Title: Intracellular Glycerol Levels Modulate the Activity of Sln1p, a Saccharomyces cerevisiae Two-component Regulator *
Article Snippet: A 1.9-kb Ava I- Bam HI restriction fragment was cloned into pUC19 (Promega Biotech) to generate the intermediate plasmid, pJF1068. .. A 2.2-kb Sal I- Xho I LEU2 fragment was subsequently ligated into the Sal I site to create pJF1070. pWT250 is a derivative of pRS424 , a 2- μ m TRP1 marked plasmid into which was cloned a 5-kb Eco RI fragment containing the GPD1 gene isolated from YEpGPD1 ( ).

Purification:

Article Title: Intracellular trafficking of nucleic acids
Article Snippet: When protein-free, purified SV40 DNA (5243 bp) was microinjected into the cytoplasm of TC7 African Green monkey kidney cells, the majority of plasmids were detected in cell nuclei by 6 – 8 h after injection by in situ hybridisation. .. In these non-dividing cells, whereas SV40 DNA localised in the nuclei with 8 h, other plasmids, including pBR322, pUC19 and pGL3-basic (Promega Corp.), did not enter the nuclei until the cells were allowed to divide [ , ].

Article Title: Host Ranges of Listeria-Specific Bacteriophages from the Turkey Processing Plant Environment in the United States ▿
Article Snippet: Phage 20422-1 DNA was purified from phage isolated following infections of L. monocytogenes DP-L862 using the Lambda miniprep phage extraction kit (Qiagen, Valencia, CA). .. The DNA was digested with NheI (New England Biolabs, Waverly, MA) and cloned into pUC19 (Promega, Madison, WI) digested with XbaI (New England Biolabs).

Lambda Miniprep:

Article Title: Host Ranges of Listeria-Specific Bacteriophages from the Turkey Processing Plant Environment in the United States ▿
Article Snippet: Phage 20422-1 DNA was purified from phage isolated following infections of L. monocytogenes DP-L862 using the Lambda miniprep phage extraction kit (Qiagen, Valencia, CA). .. The DNA was digested with NheI (New England Biolabs, Waverly, MA) and cloned into pUC19 (Promega, Madison, WI) digested with XbaI (New England Biolabs).

Sequencing:

Article Title: Intracellular trafficking of nucleic acids
Article Snippet: Paragraph title: 3.3 Sequence-specific nuclear import of plasmid DNA ... In these non-dividing cells, whereas SV40 DNA localised in the nuclei with 8 h, other plasmids, including pBR322, pUC19 and pGL3-basic (Promega Corp.), did not enter the nuclei until the cells were allowed to divide [ , ].

Article Title: The cysteine desulfurase, IscS, has a major role in in vivo Fe-S cluster formation in Escherichia coli
Article Snippet: Plasmids pPK4071 and pPK4083 were made by digesting λ430 ( ) with Nru I and cloning this 5-kb fragment containing iscRSUAhscB ( ) into the Sma I site of pUC19 ( ) and pAlter-1 (Promega), respectively. .. Plasmid pPK4344 was generated by cloning into pGem2 (Promega), a PCR product containing sequence beginning within the iscR ORF and ending within the iscA ORF (Fig. ).

Article Title: Cap-Independent Translational Enhancement of Turnip Crinkle Virus Genomic and Subgenomic RNAs
Article Snippet: .. The sgUTR-luc-3′TCV construct was made as follows: (i) the cDNA of the 1.45-kb sgRNA was PCR amplified from mutant mprNco by using one oligodeoxyribonucleotide containing the T7 promoter sequence as its 5′ half and TCV sequence from nucleotide (nt) 2607 to 2630 as its 3′ half and another oligodeoxyribonucleotide with sequence complementary to TCV nt 4054 to 4030; (ii) the PCR-amplified fragment was cloned into pUC19, and a Sna BI site was introduced at the 3′ end of the TCV CP coding region; (iii) the resulting plasmid was cut with Nco I and Sna BI, and the CP coding region was replaced with the firefly luciferase gene ( luc ) obtained by cutting pSP-luc+ (Promega, Madison, Wis.) with Nco I and Xba I. .. All other luc -containing constructs were derived from the sgUTR-luc-3′TCV by manipulating either the 5′ or 3′ UTR region or both.

Article Title: Inhibition of Hepatitis B Virus Replication by MyD88 Involves Accelerated Degradation of Pregenomic RNA and Nuclear Retention of Pre-S/S RNAs ▿
Article Snippet: To construct HBV plasmid pHBV1.3, a terminally redundant (1.3× copy), replication-competent HBV genome (subtype adw; nucleotides 957 to 1952; GenBank accession number ) was inserted into pUC19 (Promega). pCIdA-HBV expresses all the HBV proteins, but it lacks the 5′ ɛ RNA signal, which eliminates pregenomic RNA packaging and DNA replication ( ). .. Both pCMV-HBV ( ) and pTet-HBV ( ) contain the wild-type HBV 1.1-mer overlength genomic sequence.

Article Title: Host Ranges of Listeria-Specific Bacteriophages from the Turkey Processing Plant Environment in the United States ▿
Article Snippet: Paragraph title: Nucleotide sequence determinations of genomic fragments of phages 20422-1 and 805405-1. ... The DNA was digested with NheI (New England Biolabs, Waverly, MA) and cloned into pUC19 (Promega, Madison, WI) digested with XbaI (New England Biolabs).

Article Title: Tn5 Transposase with an Altered Specificity for Transposon Ends
Article Snippet: The full-length Eco RI- Kpn I transposon fragment was ligated into pUC19 to create pRZ1495/pUC. pRZ1495/pUC was digested with Not I and Afl II, filled in with dNTPs and T4 DNA polymerase (Promega), and religated to form pRZΔ1495/pUC. .. The two OE binding sites of the transposon were then mutated into the sequence IE12A following the manufacturer’s protocol for the Altered Sites system (Promega).

Article Title: Intracellular Glycerol Levels Modulate the Activity of Sln1p, a Saccharomyces cerevisiae Two-component Regulator *
Article Snippet: The presence and orientation of Myc sequences was confirmed by DNA sequence analysis. .. A 1.9-kb Ava I- Bam HI restriction fragment was cloned into pUC19 (Promega Biotech) to generate the intermediate plasmid, pJF1068.

Positron Emission Tomography:

Article Title: The Cell Cycle-Specific Growth-Inhibitory Factor Produced by Actinobacillus actinomycetemcomitans Is a Cytolethal Distending Toxin
Article Snippet: .. Manipulation of DNA in E. coli was carried out with pUC19 , pGEM-T Easy (Promega, Madison, Wis.), or pET-28a(+) (Novagen, Madison, Wis.). .. HeLa cells (ATCC CCL2) were grown in Eagle’s minimal essential medium (Nissui) supplemented with 10% fetal bovine serum at 37°C and in a 5% CO2 –95% air atmosphere.

Nested PCR:

Article Title: Quantifying promoter activity during the developmental cycle of Chlamydia trachomatis
Article Snippet: The 5′ end of the gfp transcript, which is initiated by P3, was then amplified with random-primer reverse transcription and nested PCR using gfp specific primers with the provided primers in the kit. .. PCR products were then inserted into pUC19 (Promega, Madison, WI) for DNA sequencing.

Rapid Amplification of cDNA Ends:

Article Title: Quantifying promoter activity during the developmental cycle of Chlamydia trachomatis
Article Snippet: Paragraph title: 5′ Rapid amplification of cDNA ends (RACE) ... PCR products were then inserted into pUC19 (Promega, Madison, WI) for DNA sequencing.

Antiviral Assay:

Article Title: Intracellular Glycerol Levels Modulate the Activity of Sln1p, a Saccharomyces cerevisiae Two-component Regulator *
Article Snippet: .. A 1.9-kb Ava I- Bam HI restriction fragment was cloned into pUC19 (Promega Biotech) to generate the intermediate plasmid, pJF1068. .. A deletion of 268 base pairs was introduced into the GPD1 open reading frame on pJF1068 by Sal I digestion and ligation.

Plasmid Preparation:

Article Title: Intracellular trafficking of nucleic acids
Article Snippet: Paragraph title: 3.3 Sequence-specific nuclear import of plasmid DNA ... In these non-dividing cells, whereas SV40 DNA localised in the nuclei with 8 h, other plasmids, including pBR322, pUC19 and pGL3-basic (Promega Corp.), did not enter the nuclei until the cells were allowed to divide [ , ].

Article Title: The cysteine desulfurase, IscS, has a major role in in vivo Fe-S cluster formation in Escherichia coli
Article Snippet: Paragraph title: Plasmid Construction. ... Plasmids pPK4071 and pPK4083 were made by digesting λ430 ( ) with Nru I and cloning this 5-kb fragment containing iscRSUAhscB ( ) into the Sma I site of pUC19 ( ) and pAlter-1 (Promega), respectively.

Article Title: Cap-Independent Translational Enhancement of Turnip Crinkle Virus Genomic and Subgenomic RNAs
Article Snippet: .. The sgUTR-luc-3′TCV construct was made as follows: (i) the cDNA of the 1.45-kb sgRNA was PCR amplified from mutant mprNco by using one oligodeoxyribonucleotide containing the T7 promoter sequence as its 5′ half and TCV sequence from nucleotide (nt) 2607 to 2630 as its 3′ half and another oligodeoxyribonucleotide with sequence complementary to TCV nt 4054 to 4030; (ii) the PCR-amplified fragment was cloned into pUC19, and a Sna BI site was introduced at the 3′ end of the TCV CP coding region; (iii) the resulting plasmid was cut with Nco I and Sna BI, and the CP coding region was replaced with the firefly luciferase gene ( luc ) obtained by cutting pSP-luc+ (Promega, Madison, Wis.) with Nco I and Xba I. .. All other luc -containing constructs were derived from the sgUTR-luc-3′TCV by manipulating either the 5′ or 3′ UTR region or both.

Article Title: Inhibition of Hepatitis B Virus Replication by MyD88 Involves Accelerated Degradation of Pregenomic RNA and Nuclear Retention of Pre-S/S RNAs ▿
Article Snippet: .. To construct HBV plasmid pHBV1.3, a terminally redundant (1.3× copy), replication-competent HBV genome (subtype adw; nucleotides 957 to 1952; GenBank accession number ) was inserted into pUC19 (Promega). pCIdA-HBV expresses all the HBV proteins, but it lacks the 5′ ɛ RNA signal, which eliminates pregenomic RNA packaging and DNA replication ( ). .. Both pCMV-HBV ( ) and pTet-HBV ( ) contain the wild-type HBV 1.1-mer overlength genomic sequence.

Article Title: Cloning, Characterization, Controlled Overexpression, and Inactivation of the Major Tributyrin Esterase Gene of Lactococcus lactis
Article Snippet: .. The plasmid vectors used in the cloning experiments with E. coli were pUC18 and pUC19 ( ) and pGEM-5Zf(+) (Promega Corp.). ..

Article Title: Measurement of Biologically Available Naphthalene in Gas and Aqueous Phases by Use of a Pseudomonas putida Biosensor
Article Snippet: This strain still contains the NAH plasmid and is therefore capable of using naphthalene as a sole carbon and energy source. .. The plasmids used during this study included pUC19 , for general subclonings; pGEM-T-Easy (Promega Corp.), for cloning of DNA fragments generated in PCRs; pUC18Not , for providing Not I symmetrical restriction sites; and PCK218 , serving as the mini-Tn 5 vehicle to deliver the nahR-nahG ′:: luxAB construct to the chromosome of P. putida pPG7.

Article Title: Tn5 Transposase with an Altered Specificity for Transposon Ends
Article Snippet: The papillation vector, pRZ9904 (IE12A/IE12A), was constructed in the following manner. pRZ1495 ( ) was digested to completion with Kpn I and partially digested with Eco RI. .. The full-length Eco RI- Kpn I transposon fragment was ligated into pUC19 to create pRZ1495/pUC. pRZ1495/pUC was digested with Not I and Afl II, filled in with dNTPs and T4 DNA polymerase (Promega), and religated to form pRZΔ1495/pUC.

Article Title: Intracellular Glycerol Levels Modulate the Activity of Sln1p, a Saccharomyces cerevisiae Two-component Regulator *
Article Snippet: .. A 1.9-kb Ava I- Bam HI restriction fragment was cloned into pUC19 (Promega Biotech) to generate the intermediate plasmid, pJF1068. .. A deletion of 268 base pairs was introduced into the GPD1 open reading frame on pJF1068 by Sal I digestion and ligation.

Functional Assay:

Article Title: Insights into functional and evolutionary analysis of carbaryl metabolic pathway from Pseudomonas sp. strain C5pp
Article Snippet: Paragraph title: Construction and functional screening of Pseudomonas sp. strain C5pp genomic fosmid library ... One of the randomly chosen clone designated as G1 was digested with Bam HI and all six fragments were sub-cloned into pUC19 and partially sequenced using universal M13 forward and reverse primers (M13F, 5′-GTTTTCCCAGTCACGAC-3′ and M13R, 5′-CAGGAAACAGCTATGAC-3′, Promega, USA) after cloning it into pUC19 at Bam HI site.

Binding Assay:

Article Title: Tn5 Transposase with an Altered Specificity for Transposon Ends
Article Snippet: The full-length Eco RI- Kpn I transposon fragment was ligated into pUC19 to create pRZ1495/pUC. pRZ1495/pUC was digested with Not I and Afl II, filled in with dNTPs and T4 DNA polymerase (Promega), and religated to form pRZΔ1495/pUC. .. The two OE binding sites of the transposon were then mutated into the sequence IE12A following the manufacturer’s protocol for the Altered Sites system (Promega).

In Situ:

Article Title: Intracellular trafficking of nucleic acids
Article Snippet: When protein-free, purified SV40 DNA (5243 bp) was microinjected into the cytoplasm of TC7 African Green monkey kidney cells, the majority of plasmids were detected in cell nuclei by 6 – 8 h after injection by in situ hybridisation. .. In these non-dividing cells, whereas SV40 DNA localised in the nuclei with 8 h, other plasmids, including pBR322, pUC19 and pGL3-basic (Promega Corp.), did not enter the nuclei until the cells were allowed to divide [ , ].

Protein Binding:

Article Title: Inhibition of Hepatitis B Virus Replication by MyD88 Involves Accelerated Degradation of Pregenomic RNA and Nuclear Retention of Pre-S/S RNAs ▿
Article Snippet: To construct HBV plasmid pHBV1.3, a terminally redundant (1.3× copy), replication-competent HBV genome (subtype adw; nucleotides 957 to 1952; GenBank accession number ) was inserted into pUC19 (Promega). pCIdA-HBV expresses all the HBV proteins, but it lacks the 5′ ɛ RNA signal, which eliminates pregenomic RNA packaging and DNA replication ( ). .. For these plasmids, the synthesis of the pregenomic RNA is driven by the cytomegalovirus (CMV) promoter and the Tet promoter, respectively. pCMV-HBV M2 is a derivative of pCMV-HBV in which the La protein-binding sites have been mutated ( ). pCMV-HBVΔPRE is a PRE deletion mutant of pCMV-HBV ( ).

DNA Purification:

Article Title: Insights into functional and evolutionary analysis of carbaryl metabolic pathway from Pseudomonas sp. strain C5pp
Article Snippet: Fosmid DNA was isolated by Fosmid Max DNA purification kit (FMAX 046, USA) and restriction digested with Bam HI (~12, 8, 5, 2.8, 2.5 and 1.8 kb) or Not I (20, 11, 5, 4.8, 2.7 and 2.3 kb). .. One of the randomly chosen clone designated as G1 was digested with Bam HI and all six fragments were sub-cloned into pUC19 and partially sequenced using universal M13 forward and reverse primers (M13F, 5′-GTTTTCCCAGTCACGAC-3′ and M13R, 5′-CAGGAAACAGCTATGAC-3′, Promega, USA) after cloning it into pUC19 at Bam HI site.

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  • 91
    Promega puc19
    Msc I partial digestion of talC-Bam HI fragment cloned in <t>pUC19.</t> 2 μg plasmid DNA was digested with 5 u Msc I enzyme. The digestion times are given in minutes above the gel. M, 100 bp DNA ladder makers.
    Puc19, supplied by Promega, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/puc19/product/Promega
    Average 91 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    puc19 - by Bioz Stars, 2020-02
    91/100 stars
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    94
    Promega plasmid puc19
    Steady-state fluorescence intensity ( A ) and fluorescence anisotropy ( B ) of <t>BOBO-1/pUC19</t> complexes (30 mM Tris/HCl, pH 7.4) with different dye/base ratios, at several incubation times: 10 min (⋄), 30 min (○); 60 min (*); and 90 min (•). [DNA] = 0.006 μ M.
    Plasmid Puc19, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plasmid puc19/product/Promega
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    Price from $9.99 to $1999.99
    plasmid puc19 - by Bioz Stars, 2020-02
    94/100 stars
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    81
    Promega puc19 plasmid dna
    Differences between G signals complementary to unmethylated C residues, N 4 -methylcytosine residues and 5-methylcytosine residues using the DYEnamic ET Terminator Kit. A1: untreated <t>pUC19</t> <t>DNA;</t> A2: M.BamHI N 4 -cytosine methylated pUC19DNA; A3: trace difference between A2 and A1. B1: untreated pUC19DNA; B2: M.HaeIII 5-cytosine methylated pUC19 DNA; B3: trace difference between B2 and B1. The G residues complementary to the methylated cytosines are boxed. N 4 -methylcytosine results in an increase in the complementary G signal, whereas 5-methylcytosine results in a decrease in the complementary G signal.
    Puc19 Plasmid Dna, supplied by Promega, used in various techniques. Bioz Stars score: 81/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/puc19 plasmid dna/product/Promega
    Average 81 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    Msc I partial digestion of talC-Bam HI fragment cloned in pUC19. 2 μg plasmid DNA was digested with 5 u Msc I enzyme. The digestion times are given in minutes above the gel. M, 100 bp DNA ladder makers.

    Journal: Scientific Reports

    Article Title: Rapid and efficient genome-wide characterization of Xanthomonas TAL effector genes

    doi: 10.1038/srep13162

    Figure Lengend Snippet: Msc I partial digestion of talC-Bam HI fragment cloned in pUC19. 2 μg plasmid DNA was digested with 5 u Msc I enzyme. The digestion times are given in minutes above the gel. M, 100 bp DNA ladder makers.

    Article Snippet: The obtained Bam HI fragments were ligated into Bam HI-cleaved pUC19 vector as follow: 60 ng of purified DNA fragments, 100 ng of pUC19, 2 μl of ligation buffer, and 5 u of T4 ligase (Promega) were mixed; the final reaction volume was adjusted to 20 μl with water.

    Techniques: Clone Assay, Plasmid Preparation

    Autoradiograph of an SDS–12% polyacrylamide gel containing [ 35 S]methionine-labelled products of in vitro transcription-translation. Lanes: 1, pGEM-T Easy; 2, pTK3251, containing the A. actinomycetemcomitans cdtA gene; 3, pTK3252, containing the A. actinomycetemcomitans cdtB gene; 4, pTK3253, containing the A. actinomycetemcomitans cdtC gene; 5, pTK3022, containing the entire cdtABC gene cluster; 6, pUC19. The CDT activities of sterile sonic lysates from the recombinant strains are indicated at the bottom. Radiolabelled bands marked by #, ∗, and + are putative gene products of cdtA , - B , and - C , respectively.

    Journal: Infection and Immunity

    Article Title: The Cell Cycle-Specific Growth-Inhibitory Factor Produced by Actinobacillus actinomycetemcomitans Is a Cytolethal Distending Toxin

    doi:

    Figure Lengend Snippet: Autoradiograph of an SDS–12% polyacrylamide gel containing [ 35 S]methionine-labelled products of in vitro transcription-translation. Lanes: 1, pGEM-T Easy; 2, pTK3251, containing the A. actinomycetemcomitans cdtA gene; 3, pTK3252, containing the A. actinomycetemcomitans cdtB gene; 4, pTK3253, containing the A. actinomycetemcomitans cdtC gene; 5, pTK3022, containing the entire cdtABC gene cluster; 6, pUC19. The CDT activities of sterile sonic lysates from the recombinant strains are indicated at the bottom. Radiolabelled bands marked by #, ∗, and + are putative gene products of cdtA , - B , and - C , respectively.

    Article Snippet: Manipulation of DNA in E. coli was carried out with pUC19 , pGEM-T Easy (Promega, Madison, Wis.), or pET-28a(+) (Novagen, Madison, Wis.).

    Techniques: Autoradiography, In Vitro, Recombinant

    Steady-state fluorescence intensity ( A ) and fluorescence anisotropy ( B ) of BOBO-1/pUC19 complexes (30 mM Tris/HCl, pH 7.4) with different dye/base ratios, at several incubation times: 10 min (⋄), 30 min (○); 60 min (*); and 90 min (•). [DNA] = 0.006 μ M.

    Journal: Biophysical Journal

    Article Title: Characterization of DNA/Lipid Complexes by Fluorescence Resonance Energy Transfer

    doi:

    Figure Lengend Snippet: Steady-state fluorescence intensity ( A ) and fluorescence anisotropy ( B ) of BOBO-1/pUC19 complexes (30 mM Tris/HCl, pH 7.4) with different dye/base ratios, at several incubation times: 10 min (⋄), 30 min (○); 60 min (*); and 90 min (•). [DNA] = 0.006 μ M.

    Article Snippet: The plasmid pUC19 (2690 bp) was purchased from Promega (Madison, WI).

    Techniques: Fluorescence, Incubation

    Mean diameter of pUC19 without (○), and with (•), BOBO-1 at d / b = 0.01, with cationic liposomes (DOTAP) at several charge ratio (+/−). [DNA] = 0.007 μ M. The line is a mere guide to the eye.

    Journal: Biophysical Journal

    Article Title: Characterization of DNA/Lipid Complexes by Fluorescence Resonance Energy Transfer

    doi:

    Figure Lengend Snippet: Mean diameter of pUC19 without (○), and with (•), BOBO-1 at d / b = 0.01, with cationic liposomes (DOTAP) at several charge ratio (+/−). [DNA] = 0.007 μ M. The line is a mere guide to the eye.

    Article Snippet: The plasmid pUC19 (2690 bp) was purchased from Promega (Madison, WI).

    Techniques:

    Schematic representation of the lipoplexes multilamellar structure with the fluorescent probes within the DNA and the lipid. ( A ) Acceptor ( a ) on the DNA (EtBr) and donor ( d ) on the lipid (DPH-PC and BODIPY-PC). This arrangement was used for DOTAP/pUC19 charge ratio (+/−) = 0.5. ( B ) Acceptor on the lipid (BODIPY-PC) and donor on the DNA (BOBO-1). This arrangement was used for DOTAP/pUC19 charge ratio (+/−) = 0.5, 2, and 4.

    Journal: Biophysical Journal

    Article Title: Characterization of DNA/Lipid Complexes by Fluorescence Resonance Energy Transfer

    doi:

    Figure Lengend Snippet: Schematic representation of the lipoplexes multilamellar structure with the fluorescent probes within the DNA and the lipid. ( A ) Acceptor ( a ) on the DNA (EtBr) and donor ( d ) on the lipid (DPH-PC and BODIPY-PC). This arrangement was used for DOTAP/pUC19 charge ratio (+/−) = 0.5. ( B ) Acceptor on the lipid (BODIPY-PC) and donor on the DNA (BOBO-1). This arrangement was used for DOTAP/pUC19 charge ratio (+/−) = 0.5, 2, and 4.

    Article Snippet: The plasmid pUC19 (2690 bp) was purchased from Promega (Madison, WI).

    Techniques:

    Electrophoretic profile of BOBO-1/pUC19 complexes (30 mM Tris/HCl, pH 7.4) at dye:DNA base ( d / b ) values: 0 ( lane 1 ), 0.2 ( lane 2 ), 0.167 ( lane 3), 0.09 ( lane 4 ), 0.06 ( lane 5 ), 0.03 ( lane 6 ), and 0.01 ( lane 7 ), after 30 min incubation at room temperature. [DNA] = 0.03 μ M.

    Journal: Biophysical Journal

    Article Title: Characterization of DNA/Lipid Complexes by Fluorescence Resonance Energy Transfer

    doi:

    Figure Lengend Snippet: Electrophoretic profile of BOBO-1/pUC19 complexes (30 mM Tris/HCl, pH 7.4) at dye:DNA base ( d / b ) values: 0 ( lane 1 ), 0.2 ( lane 2 ), 0.167 ( lane 3), 0.09 ( lane 4 ), 0.06 ( lane 5 ), 0.03 ( lane 6 ), and 0.01 ( lane 7 ), after 30 min incubation at room temperature. [DNA] = 0.03 μ M.

    Article Snippet: The plasmid pUC19 (2690 bp) was purchased from Promega (Madison, WI).

    Techniques: Incubation

    Differences between G signals complementary to unmethylated C residues, N 4 -methylcytosine residues and 5-methylcytosine residues using the DYEnamic ET Terminator Kit. A1: untreated pUC19 DNA; A2: M.BamHI N 4 -cytosine methylated pUC19DNA; A3: trace difference between A2 and A1. B1: untreated pUC19DNA; B2: M.HaeIII 5-cytosine methylated pUC19 DNA; B3: trace difference between B2 and B1. The G residues complementary to the methylated cytosines are boxed. N 4 -methylcytosine results in an increase in the complementary G signal, whereas 5-methylcytosine results in a decrease in the complementary G signal.

    Journal: Nucleic Acids Research

    Article Title: Direct detection of methylation in genomic DNA

    doi: 10.1093/nar/gni121

    Figure Lengend Snippet: Differences between G signals complementary to unmethylated C residues, N 4 -methylcytosine residues and 5-methylcytosine residues using the DYEnamic ET Terminator Kit. A1: untreated pUC19 DNA; A2: M.BamHI N 4 -cytosine methylated pUC19DNA; A3: trace difference between A2 and A1. B1: untreated pUC19DNA; B2: M.HaeIII 5-cytosine methylated pUC19 DNA; B3: trace difference between B2 and B1. The G residues complementary to the methylated cytosines are boxed. N 4 -methylcytosine results in an increase in the complementary G signal, whereas 5-methylcytosine results in a decrease in the complementary G signal.

    Article Snippet: Chromosomal DNA of H.pylori strain 1061 was isolated as described earlier ( ). pUC19 plasmid DNA was isolated from E.coli DH5α using the Wizard plus SV miniprep kit (Promega).

    Techniques: Methylation