ndei  (Millipore)


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    Nde I
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    ndei-ro
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    Structured Review

    Millipore ndei
    Cloning, expression, and purification of rPCNA. (A) Specific PCR of <t>PCNA.</t> Lane 1, 1-kb ladder; lanes 2 and 3, amplified PCR product at 882 bp. (B) Clone confirmation in pTZ57R/T. Lane 1, 1-kb ladder; lane 2; <t>NdeI-</t> and BamHI-digested pTZ57R/T-PCNA. (C)

    https://www.bioz.com/result/ndei/product/Millipore
    Average 99 stars, based on 1180 article reviews
    Price from $9.99 to $1999.99
    ndei - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Characterization of the Proliferating Cell Nuclear Antigen of Leishmania donovani Clinical Isolates and Its Association with Antimony Resistance"

    Article Title: Characterization of the Proliferating Cell Nuclear Antigen of Leishmania donovani Clinical Isolates and Its Association with Antimony Resistance

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.01847-13

    Cloning, expression, and purification of rPCNA. (A) Specific PCR of PCNA. Lane 1, 1-kb ladder; lanes 2 and 3, amplified PCR product at 882 bp. (B) Clone confirmation in pTZ57R/T. Lane 1, 1-kb ladder; lane 2; NdeI- and BamHI-digested pTZ57R/T-PCNA. (C)
    Figure Legend Snippet: Cloning, expression, and purification of rPCNA. (A) Specific PCR of PCNA. Lane 1, 1-kb ladder; lanes 2 and 3, amplified PCR product at 882 bp. (B) Clone confirmation in pTZ57R/T. Lane 1, 1-kb ladder; lane 2; NdeI- and BamHI-digested pTZ57R/T-PCNA. (C)

    Techniques Used: Clone Assay, Expressing, Purification, Polymerase Chain Reaction, Amplification

    2) Product Images from "Nuclear MxA proteins form a complex with influenza virus NP and inhibit the transcription of the engineered influenza virus genome"

    Article Title: Nuclear MxA proteins form a complex with influenza virus NP and inhibit the transcription of the engineered influenza virus genome

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkh192

    The effect of transiently expressed nuclear MxA proteins. ( A ) The effect of nuclear MxA proteins in Swiss3T3 cells. The cells were transfected with pHMP1-vNS-Luc (0.2 µg), pSEAP2-control (0.1 µg), plasmids encoding three RNA polymerase subunits and NP (0.05 µg of each plasmid) and either parental vector, pVP16-MxA or pHMG-TMxA (0.3 µg each). Luciferase activity was determined and shown as described in Materials and Methods. Error bars represent standard deviation (n = 3). ( B ) Dose-dependent effect of wild-type MxA and VP16-MxA. The same procedure for (A) was carried out in the presence of increasing amounts of wild-type MxA or VP16-MxA. Error bars represent standard deviation (n = 3). ( C ) The effect of C-terminal (MxAΔC) and internal (MxAΔM) deletion VP16-MxA mutant proteins on reporter gene expression. The same procedure for (A) was carried out with mutant MxA proteins. To generate a VP16-MxA deletion mutant (MxAΔC) for expression of MxA lacking its C-terminal region (362–662), we amplified a fragment by PCR with specific primers, 5′-GGCATCCATATGGTTGTTTCCGAAGTGGACATCGCA-3′ and 5′-CGCGGATCCTTAACCATACTTTTGTAGCTCCTCTGT-3′ and pVP16-MxA as template. To generate a VP16-MxA deletion mutant (MxAΔM) lacking its internal region (362–573), a fragment was amplified by PCR with primers, 5′-GGCATCCATATGGTTGTTTCCGAAGTGGACATC GCA-3′ and 5′-CGCGGATCCTTAACCGGGGAACTGGGCAAGCCGGCG-3′ and pCHA-MxAΔM plasmid as a template. pCHA-MxAΔM was derived from previously constructed plasmid, pCHA-MxA by removal of an internal part of MxA by digestion with SalI (TOYOBO) and NcoI (TOYOBO) restriction enzymes. The main part of the plasmid was blunted with Klenow fragment and self-ligated. MxA fragments thus prepared were digested with NdeI, blunted with Klenow fragment and then digested with BamHI. These fragments were cloned into pVP16 plasmid digested with EcoRI followed by Klenow treatment and subsequent digestion with BamHI. Error bars represent standard deviation (n = 3).
    Figure Legend Snippet: The effect of transiently expressed nuclear MxA proteins. ( A ) The effect of nuclear MxA proteins in Swiss3T3 cells. The cells were transfected with pHMP1-vNS-Luc (0.2 µg), pSEAP2-control (0.1 µg), plasmids encoding three RNA polymerase subunits and NP (0.05 µg of each plasmid) and either parental vector, pVP16-MxA or pHMG-TMxA (0.3 µg each). Luciferase activity was determined and shown as described in Materials and Methods. Error bars represent standard deviation (n = 3). ( B ) Dose-dependent effect of wild-type MxA and VP16-MxA. The same procedure for (A) was carried out in the presence of increasing amounts of wild-type MxA or VP16-MxA. Error bars represent standard deviation (n = 3). ( C ) The effect of C-terminal (MxAΔC) and internal (MxAΔM) deletion VP16-MxA mutant proteins on reporter gene expression. The same procedure for (A) was carried out with mutant MxA proteins. To generate a VP16-MxA deletion mutant (MxAΔC) for expression of MxA lacking its C-terminal region (362–662), we amplified a fragment by PCR with specific primers, 5′-GGCATCCATATGGTTGTTTCCGAAGTGGACATCGCA-3′ and 5′-CGCGGATCCTTAACCATACTTTTGTAGCTCCTCTGT-3′ and pVP16-MxA as template. To generate a VP16-MxA deletion mutant (MxAΔM) lacking its internal region (362–573), a fragment was amplified by PCR with primers, 5′-GGCATCCATATGGTTGTTTCCGAAGTGGACATC GCA-3′ and 5′-CGCGGATCCTTAACCGGGGAACTGGGCAAGCCGGCG-3′ and pCHA-MxAΔM plasmid as a template. pCHA-MxAΔM was derived from previously constructed plasmid, pCHA-MxA by removal of an internal part of MxA by digestion with SalI (TOYOBO) and NcoI (TOYOBO) restriction enzymes. The main part of the plasmid was blunted with Klenow fragment and self-ligated. MxA fragments thus prepared were digested with NdeI, blunted with Klenow fragment and then digested with BamHI. These fragments were cloned into pVP16 plasmid digested with EcoRI followed by Klenow treatment and subsequent digestion with BamHI. Error bars represent standard deviation (n = 3).

    Techniques Used: Transfection, Plasmid Preparation, Luciferase, Activity Assay, Standard Deviation, Mutagenesis, Expressing, Amplification, Polymerase Chain Reaction, Derivative Assay, Construct, Clone Assay

    Related Articles

    Clone Assay:

    Article Title: New ex vivo reporter assay system reveals that ? factors of an unculturable pathogen control gene regulation involved in the host switching between insects and plants
    Article Snippet: .. The PCR product was digested with Nde I and Xho I restriction enzymes for rpoD , or Nde I and Hin dIII for fliA , and then cloned into the pET-30a vectors (Novagen, Madison, WI) through the same sites. .. Next, the promoter region of OY-M rrnB (300 bp upstream of the gene), luciferase , and gfp were separately amplified by PCR using the primer sets described in Table S3 and KOD DNA polymerase.

    Article Title: Genetic and Physiological Characteristics of a Novel Marine Propylene-Assimilating Halieaceae Bacterium Isolated from Seawater and the Diversity of Its Alkene and Epoxide Metabolism Genes
    Article Snippet: .. The amplified fragment was digested with Nde I and Bam HI and cloned into pET15b(+) (Novagen, Madison, WI) to obtain pSUPE-15E. pSUPE-15E clones were transformed into E. coli BL21(DE3) and cultivated in LB liquid medium containing 100 μg mL−1 ampicillin at 28°C for 6 h. They were cultured for a further 16 h after the addition of IPTG (1 mM). .. The culture broth of E. coli BL21(DE3) harboring pSUPE-15E was harvested by centrifugation at 8,000 rpm and washed once with 50 mM PPB (50 mM potassium dihydrogen phosphate and 50 mM dipotassium hydrogen phosphate, pH 8.0).

    Amplification:

    Article Title: Sequence conservation of Plasmodium vivax glutamate dehydrogenase among Korean isolates and its application in seroepidemiology
    Article Snippet: .. The amplified PCR product was digested with Nde I and Xho I, purified with a QIAquick Gel Extraction Kit, and integrated into the Nde I and Xho I sites of the pET28b expression vector (Novagen). .. The resulting plasmid (named pVKtype3) was subsequently used for the expression of a pGDH-(His)6 fusion protein in E. coli .

    Article Title: Genetic and Physiological Characteristics of a Novel Marine Propylene-Assimilating Halieaceae Bacterium Isolated from Seawater and the Diversity of Its Alkene and Epoxide Metabolism Genes
    Article Snippet: .. The amplified fragment was digested with Nde I and Bam HI and cloned into pET15b(+) (Novagen, Madison, WI) to obtain pSUPE-15E. pSUPE-15E clones were transformed into E. coli BL21(DE3) and cultivated in LB liquid medium containing 100 μg mL−1 ampicillin at 28°C for 6 h. They were cultured for a further 16 h after the addition of IPTG (1 mM). .. The culture broth of E. coli BL21(DE3) harboring pSUPE-15E was harvested by centrifugation at 8,000 rpm and washed once with 50 mM PPB (50 mM potassium dihydrogen phosphate and 50 mM dipotassium hydrogen phosphate, pH 8.0).

    Article Title: Protective Immunization with a Novel Membrane Protein of Plasmodium yoelii-Infected Erythrocytes
    Article Snippet: .. The amplified fragments were gel purified, digested with Nde I and Xho I, and ligated into Nde I/ Xho I-digested pET-15b (Novagen, Madison, Wis.). ..

    Cell Culture:

    Article Title: Genetic and Physiological Characteristics of a Novel Marine Propylene-Assimilating Halieaceae Bacterium Isolated from Seawater and the Diversity of Its Alkene and Epoxide Metabolism Genes
    Article Snippet: .. The amplified fragment was digested with Nde I and Bam HI and cloned into pET15b(+) (Novagen, Madison, WI) to obtain pSUPE-15E. pSUPE-15E clones were transformed into E. coli BL21(DE3) and cultivated in LB liquid medium containing 100 μg mL−1 ampicillin at 28°C for 6 h. They were cultured for a further 16 h after the addition of IPTG (1 mM). .. The culture broth of E. coli BL21(DE3) harboring pSUPE-15E was harvested by centrifugation at 8,000 rpm and washed once with 50 mM PPB (50 mM potassium dihydrogen phosphate and 50 mM dipotassium hydrogen phosphate, pH 8.0).

    Purification:

    Article Title: Molecular Cloning and Characterization of P4 Nuclease from Leishmania infantum
    Article Snippet: .. Expression and Purification The pGEM vector containing Li-P4 gene was digested with Nde I and Xho I and the insert was purified, subcloned into the Nde I - Xho I digested pET28a (Novagen) expression vector and transformed into the E. coli BL21 (Novagen). .. The bacteria containing pET28a-Li-P4 was cultured in LB broth medium and grown until OD = 0.5.

    Article Title: Sequence conservation of Plasmodium vivax glutamate dehydrogenase among Korean isolates and its application in seroepidemiology
    Article Snippet: .. The amplified PCR product was digested with Nde I and Xho I, purified with a QIAquick Gel Extraction Kit, and integrated into the Nde I and Xho I sites of the pET28b expression vector (Novagen). .. The resulting plasmid (named pVKtype3) was subsequently used for the expression of a pGDH-(His)6 fusion protein in E. coli .

    Article Title: Protective Immunization with a Novel Membrane Protein of Plasmodium yoelii-Infected Erythrocytes
    Article Snippet: .. The amplified fragments were gel purified, digested with Nde I and Xho I, and ligated into Nde I/ Xho I-digested pET-15b (Novagen, Madison, Wis.). ..

    Positron Emission Tomography:

    Article Title: New ex vivo reporter assay system reveals that ? factors of an unculturable pathogen control gene regulation involved in the host switching between insects and plants
    Article Snippet: .. The PCR product was digested with Nde I and Xho I restriction enzymes for rpoD , or Nde I and Hin dIII for fliA , and then cloned into the pET-30a vectors (Novagen, Madison, WI) through the same sites. .. Next, the promoter region of OY-M rrnB (300 bp upstream of the gene), luciferase , and gfp were separately amplified by PCR using the primer sets described in Table S3 and KOD DNA polymerase.

    Article Title: Protective Immunization with a Novel Membrane Protein of Plasmodium yoelii-Infected Erythrocytes
    Article Snippet: .. The amplified fragments were gel purified, digested with Nde I and Xho I, and ligated into Nde I/ Xho I-digested pET-15b (Novagen, Madison, Wis.). ..

    Expressing:

    Article Title: Characterization of the Proliferating Cell Nuclear Antigen of Leishmania donovani Clinical Isolates and Its Association with Antimony Resistance
    Article Snippet: .. PCNA was further subcloned at the NdeI and BamHI sites in the bacterial expression vector pET28a (Novagen). .. The expression of rLdPCNA was checked in bacterial cells by transforming the PCNA-pET28a construct in the E. coli Rosetta strain.

    Article Title: Molecular Cloning and Characterization of P4 Nuclease from Leishmania infantum
    Article Snippet: .. Expression and Purification The pGEM vector containing Li-P4 gene was digested with Nde I and Xho I and the insert was purified, subcloned into the Nde I - Xho I digested pET28a (Novagen) expression vector and transformed into the E. coli BL21 (Novagen). .. The bacteria containing pET28a-Li-P4 was cultured in LB broth medium and grown until OD = 0.5.

    Article Title: Sequence conservation of Plasmodium vivax glutamate dehydrogenase among Korean isolates and its application in seroepidemiology
    Article Snippet: .. The amplified PCR product was digested with Nde I and Xho I, purified with a QIAquick Gel Extraction Kit, and integrated into the Nde I and Xho I sites of the pET28b expression vector (Novagen). .. The resulting plasmid (named pVKtype3) was subsequently used for the expression of a pGDH-(His)6 fusion protein in E. coli .

    Polymerase Chain Reaction:

    Article Title: Purification and Characterization of the RecA Protein from Neisseria gonorrhoeae
    Article Snippet: .. The PCR product was digested with Nde I and Hind III and inserted into the overproduction vector pET21a (Novagen) cut with the same restriction enzymes. .. The resulting plasmid was designated pEAW375.

    Article Title: New ex vivo reporter assay system reveals that ? factors of an unculturable pathogen control gene regulation involved in the host switching between insects and plants
    Article Snippet: .. The PCR product was digested with Nde I and Xho I restriction enzymes for rpoD , or Nde I and Hin dIII for fliA , and then cloned into the pET-30a vectors (Novagen, Madison, WI) through the same sites. .. Next, the promoter region of OY-M rrnB (300 bp upstream of the gene), luciferase , and gfp were separately amplified by PCR using the primer sets described in Table S3 and KOD DNA polymerase.

    Article Title: Sequence conservation of Plasmodium vivax glutamate dehydrogenase among Korean isolates and its application in seroepidemiology
    Article Snippet: .. The amplified PCR product was digested with Nde I and Xho I, purified with a QIAquick Gel Extraction Kit, and integrated into the Nde I and Xho I sites of the pET28b expression vector (Novagen). .. The resulting plasmid (named pVKtype3) was subsequently used for the expression of a pGDH-(His)6 fusion protein in E. coli .

    Gel Extraction:

    Article Title: Sequence conservation of Plasmodium vivax glutamate dehydrogenase among Korean isolates and its application in seroepidemiology
    Article Snippet: .. The amplified PCR product was digested with Nde I and Xho I, purified with a QIAquick Gel Extraction Kit, and integrated into the Nde I and Xho I sites of the pET28b expression vector (Novagen). .. The resulting plasmid (named pVKtype3) was subsequently used for the expression of a pGDH-(His)6 fusion protein in E. coli .

    Transformation Assay:

    Article Title: Molecular Cloning and Characterization of P4 Nuclease from Leishmania infantum
    Article Snippet: .. Expression and Purification The pGEM vector containing Li-P4 gene was digested with Nde I and Xho I and the insert was purified, subcloned into the Nde I - Xho I digested pET28a (Novagen) expression vector and transformed into the E. coli BL21 (Novagen). .. The bacteria containing pET28a-Li-P4 was cultured in LB broth medium and grown until OD = 0.5.

    Article Title: Genetic and Physiological Characteristics of a Novel Marine Propylene-Assimilating Halieaceae Bacterium Isolated from Seawater and the Diversity of Its Alkene and Epoxide Metabolism Genes
    Article Snippet: .. The amplified fragment was digested with Nde I and Bam HI and cloned into pET15b(+) (Novagen, Madison, WI) to obtain pSUPE-15E. pSUPE-15E clones were transformed into E. coli BL21(DE3) and cultivated in LB liquid medium containing 100 μg mL−1 ampicillin at 28°C for 6 h. They were cultured for a further 16 h after the addition of IPTG (1 mM). .. The culture broth of E. coli BL21(DE3) harboring pSUPE-15E was harvested by centrifugation at 8,000 rpm and washed once with 50 mM PPB (50 mM potassium dihydrogen phosphate and 50 mM dipotassium hydrogen phosphate, pH 8.0).

    Plasmid Preparation:

    Article Title: Characterization of the Proliferating Cell Nuclear Antigen of Leishmania donovani Clinical Isolates and Its Association with Antimony Resistance
    Article Snippet: .. PCNA was further subcloned at the NdeI and BamHI sites in the bacterial expression vector pET28a (Novagen). .. The expression of rLdPCNA was checked in bacterial cells by transforming the PCNA-pET28a construct in the E. coli Rosetta strain.

    Article Title: Purification and Characterization of the RecA Protein from Neisseria gonorrhoeae
    Article Snippet: .. The PCR product was digested with Nde I and Hind III and inserted into the overproduction vector pET21a (Novagen) cut with the same restriction enzymes. .. The resulting plasmid was designated pEAW375.

    Article Title: Molecular Cloning and Characterization of P4 Nuclease from Leishmania infantum
    Article Snippet: .. Expression and Purification The pGEM vector containing Li-P4 gene was digested with Nde I and Xho I and the insert was purified, subcloned into the Nde I - Xho I digested pET28a (Novagen) expression vector and transformed into the E. coli BL21 (Novagen). .. The bacteria containing pET28a-Li-P4 was cultured in LB broth medium and grown until OD = 0.5.

    Article Title: Sequence conservation of Plasmodium vivax glutamate dehydrogenase among Korean isolates and its application in seroepidemiology
    Article Snippet: .. The amplified PCR product was digested with Nde I and Xho I, purified with a QIAquick Gel Extraction Kit, and integrated into the Nde I and Xho I sites of the pET28b expression vector (Novagen). .. The resulting plasmid (named pVKtype3) was subsequently used for the expression of a pGDH-(His)6 fusion protein in E. coli .

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  • 99
    Millipore nde i
    Determination of the crossover site. (A) Sequence of the minimal attP and attB sites, with common nucleotides marked with dots. The central three nucleotides are numbered 1 to 3 in this figure. (B) In vitro intramolecular deletion reactions (30 minutes) on supercoiled plasmids with attB sites containing mutations at the central three nucleotides as designated. P×B, wild-type control. The products were digested with Xho I and Bam H1 to reveal deletion products (Figure 1D ). (C) Results of in vivo deletion reactions between attP and mutated attB sites. White colonies result from site-specific deletions and blue colonies signify absence of deletions. (D) In vitro intramolecular recombination reactions (30 minutes) with plasmid substrates containing identical changes within attP and attB at the designated positions. Parental and deletion product bands are denoted. (E) Results of in vivo recombination reactions between mutated attP and attB sites. Because G1C and G2C mutations create symmetrical cores, inversions between att sites oriented in an antiparallel configuration (G) form along with deletions. Inversion products retain the Lac + (blue) phenotype. (F) Diagram of synapsis between attP (G1C) and attB (G1C) in the standard parallel orientation generating attL and attR upon <t>DNA</t> exchange. Productive recombination between wild-type attP and attB sites containing the asymmetric GG core nucleotides only occur by this pathway. (G) Diagram of an antiparallel synapsis between attP (G1C) and attB (G1C) generating the hybrid attL * and attR * sites upon DNA exchange. (H) In vitro intramolecular recombination reactions (30 minutes) with plasmid substrates containing identical changes within attP and attB within the core nucleotides. The reaction products were digested with Bam HI, Nde I, and Sca I to reveal both deletions (doublet bands) and inversions as denoted.
    Nde I, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1096 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nde i/product/Millipore
    Average 99 stars, based on 1096 article reviews
    Price from $9.99 to $1999.99
    nde i - by Bioz Stars, 2020-09
    99/100 stars
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    85
    Millipore ndei xhoi restriction digestion
    Determination of the crossover site. (A) Sequence of the minimal attP and attB sites, with common nucleotides marked with dots. The central three nucleotides are numbered 1 to 3 in this figure. (B) In vitro intramolecular deletion reactions (30 minutes) on supercoiled plasmids with attB sites containing mutations at the central three nucleotides as designated. P×B, wild-type control. The products were digested with Xho I and Bam H1 to reveal deletion products (Figure 1D ). (C) Results of in vivo deletion reactions between attP and mutated attB sites. White colonies result from site-specific deletions and blue colonies signify absence of deletions. (D) In vitro intramolecular recombination reactions (30 minutes) with plasmid substrates containing identical changes within attP and attB at the designated positions. Parental and deletion product bands are denoted. (E) Results of in vivo recombination reactions between mutated attP and attB sites. Because G1C and G2C mutations create symmetrical cores, inversions between att sites oriented in an antiparallel configuration (G) form along with deletions. Inversion products retain the Lac + (blue) phenotype. (F) Diagram of synapsis between attP (G1C) and attB (G1C) in the standard parallel orientation generating attL and attR upon <t>DNA</t> exchange. Productive recombination between wild-type attP and attB sites containing the asymmetric GG core nucleotides only occur by this pathway. (G) Diagram of an antiparallel synapsis between attP (G1C) and attB (G1C) generating the hybrid attL * and attR * sites upon DNA exchange. (H) In vitro intramolecular recombination reactions (30 minutes) with plasmid substrates containing identical changes within attP and attB within the core nucleotides. The reaction products were digested with Bam HI, Nde I, and Sca I to reveal both deletions (doublet bands) and inversions as denoted.
    Ndei Xhoi Restriction Digestion, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ndei xhoi restriction digestion/product/Millipore
    Average 85 stars, based on 1 article reviews
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    90
    Millipore ndei noti sites
    Determination of the crossover site. (A) Sequence of the minimal attP and attB sites, with common nucleotides marked with dots. The central three nucleotides are numbered 1 to 3 in this figure. (B) In vitro intramolecular deletion reactions (30 minutes) on supercoiled plasmids with attB sites containing mutations at the central three nucleotides as designated. P×B, wild-type control. The products were digested with Xho I and Bam H1 to reveal deletion products (Figure 1D ). (C) Results of in vivo deletion reactions between attP and mutated attB sites. White colonies result from site-specific deletions and blue colonies signify absence of deletions. (D) In vitro intramolecular recombination reactions (30 minutes) with plasmid substrates containing identical changes within attP and attB at the designated positions. Parental and deletion product bands are denoted. (E) Results of in vivo recombination reactions between mutated attP and attB sites. Because G1C and G2C mutations create symmetrical cores, inversions between att sites oriented in an antiparallel configuration (G) form along with deletions. Inversion products retain the Lac + (blue) phenotype. (F) Diagram of synapsis between attP (G1C) and attB (G1C) in the standard parallel orientation generating attL and attR upon <t>DNA</t> exchange. Productive recombination between wild-type attP and attB sites containing the asymmetric GG core nucleotides only occur by this pathway. (G) Diagram of an antiparallel synapsis between attP (G1C) and attB (G1C) generating the hybrid attL * and attR * sites upon DNA exchange. (H) In vitro intramolecular recombination reactions (30 minutes) with plasmid substrates containing identical changes within attP and attB within the core nucleotides. The reaction products were digested with Bam HI, Nde I, and Sca I to reveal both deletions (doublet bands) and inversions as denoted.
    Ndei Noti Sites, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Determination of the crossover site. (A) Sequence of the minimal attP and attB sites, with common nucleotides marked with dots. The central three nucleotides are numbered 1 to 3 in this figure. (B) In vitro intramolecular deletion reactions (30 minutes) on supercoiled plasmids with attB sites containing mutations at the central three nucleotides as designated. P×B, wild-type control. The products were digested with Xho I and Bam H1 to reveal deletion products (Figure 1D ). (C) Results of in vivo deletion reactions between attP and mutated attB sites. White colonies result from site-specific deletions and blue colonies signify absence of deletions. (D) In vitro intramolecular recombination reactions (30 minutes) with plasmid substrates containing identical changes within attP and attB at the designated positions. Parental and deletion product bands are denoted. (E) Results of in vivo recombination reactions between mutated attP and attB sites. Because G1C and G2C mutations create symmetrical cores, inversions between att sites oriented in an antiparallel configuration (G) form along with deletions. Inversion products retain the Lac + (blue) phenotype. (F) Diagram of synapsis between attP (G1C) and attB (G1C) in the standard parallel orientation generating attL and attR upon DNA exchange. Productive recombination between wild-type attP and attB sites containing the asymmetric GG core nucleotides only occur by this pathway. (G) Diagram of an antiparallel synapsis between attP (G1C) and attB (G1C) generating the hybrid attL * and attR * sites upon DNA exchange. (H) In vitro intramolecular recombination reactions (30 minutes) with plasmid substrates containing identical changes within attP and attB within the core nucleotides. The reaction products were digested with Bam HI, Nde I, and Sca I to reveal both deletions (doublet bands) and inversions as denoted.

    Journal: Mobile DNA

    Article Title: The site-specific integration reaction of Listeria phage A118 integrase, a serine recombinase

    doi: 10.1186/1759-8753-4-2

    Figure Lengend Snippet: Determination of the crossover site. (A) Sequence of the minimal attP and attB sites, with common nucleotides marked with dots. The central three nucleotides are numbered 1 to 3 in this figure. (B) In vitro intramolecular deletion reactions (30 minutes) on supercoiled plasmids with attB sites containing mutations at the central three nucleotides as designated. P×B, wild-type control. The products were digested with Xho I and Bam H1 to reveal deletion products (Figure 1D ). (C) Results of in vivo deletion reactions between attP and mutated attB sites. White colonies result from site-specific deletions and blue colonies signify absence of deletions. (D) In vitro intramolecular recombination reactions (30 minutes) with plasmid substrates containing identical changes within attP and attB at the designated positions. Parental and deletion product bands are denoted. (E) Results of in vivo recombination reactions between mutated attP and attB sites. Because G1C and G2C mutations create symmetrical cores, inversions between att sites oriented in an antiparallel configuration (G) form along with deletions. Inversion products retain the Lac + (blue) phenotype. (F) Diagram of synapsis between attP (G1C) and attB (G1C) in the standard parallel orientation generating attL and attR upon DNA exchange. Productive recombination between wild-type attP and attB sites containing the asymmetric GG core nucleotides only occur by this pathway. (G) Diagram of an antiparallel synapsis between attP (G1C) and attB (G1C) generating the hybrid attL * and attR * sites upon DNA exchange. (H) In vitro intramolecular recombination reactions (30 minutes) with plasmid substrates containing identical changes within attP and attB within the core nucleotides. The reaction products were digested with Bam HI, Nde I, and Sca I to reveal both deletions (doublet bands) and inversions as denoted.

    Article Snippet: The A118 integrase coding sequence was amplified by PCR from the phage DNA and cloned between Nde I and Bam HI sites of pET11a and pET15b (EMD Millipore Billerica, MA USA) to give pRJ2186 and pRJ2184, respectively. pRJ2823 contains integrase residues 158 to 452 (C-terminus) in pET15b.

    Techniques: Sequencing, In Vitro, In Vivo, Plasmid Preparation