prc cmv ha vhl expression construct  (New England Biolabs)


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    Structured Review

    New England Biolabs prc cmv ha vhl expression construct
    MT1-MMP overexpression in RCC cells lacking <t>VHL.</t> Total RNA and protein were isolated from WT8 and <t>pRc-9</t> cells. (A) Northern blot analysis of MT1-MMP mRNA expression and western blot analysis of VHL protein expression in RCC cell lines. RNA levels were
    Prc Cmv Ha Vhl Expression Construct, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 85/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/prc cmv ha vhl expression construct/product/New England Biolabs
    Average 85 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    prc cmv ha vhl expression construct - by Bioz Stars, 2020-04
    85/100 stars

    Images

    1) Product Images from "Identification of Membrane Type-1 Matrix Metalloproteinase as a Target of Hypoxia-Inducible Factor-2? in von Hippel Lindau Renal Cell Carcinoma"

    Article Title: Identification of Membrane Type-1 Matrix Metalloproteinase as a Target of Hypoxia-Inducible Factor-2? in von Hippel Lindau Renal Cell Carcinoma

    Journal: Oncogene

    doi: 10.1038/sj.onc.1208305

    MT1-MMP overexpression in RCC cells lacking VHL. Total RNA and protein were isolated from WT8 and pRc-9 cells. (A) Northern blot analysis of MT1-MMP mRNA expression and western blot analysis of VHL protein expression in RCC cell lines. RNA levels were
    Figure Legend Snippet: MT1-MMP overexpression in RCC cells lacking VHL. Total RNA and protein were isolated from WT8 and pRc-9 cells. (A) Northern blot analysis of MT1-MMP mRNA expression and western blot analysis of VHL protein expression in RCC cell lines. RNA levels were

    Techniques Used: Over Expression, Isolation, Northern Blot, Expressing, Western Blot

    2) Product Images from "Identification of Membrane Type-1 Matrix Metalloproteinase as a Target of Hypoxia-Inducible Factor-2? in von Hippel Lindau Renal Cell Carcinoma"

    Article Title: Identification of Membrane Type-1 Matrix Metalloproteinase as a Target of Hypoxia-Inducible Factor-2? in von Hippel Lindau Renal Cell Carcinoma

    Journal: Oncogene

    doi: 10.1038/sj.onc.1208305

    MT1-MMP overexpression in RCC cells lacking VHL. Total RNA and protein were isolated from WT8 and pRc-9 cells. (A) Northern blot analysis of MT1-MMP mRNA expression and western blot analysis of VHL protein expression in RCC cell lines. RNA levels were
    Figure Legend Snippet: MT1-MMP overexpression in RCC cells lacking VHL. Total RNA and protein were isolated from WT8 and pRc-9 cells. (A) Northern blot analysis of MT1-MMP mRNA expression and western blot analysis of VHL protein expression in RCC cell lines. RNA levels were

    Techniques Used: Over Expression, Isolation, Northern Blot, Expressing, Western Blot

    3) Product Images from "Identification of Membrane Type-1 Matrix Metalloproteinase as a Target of Hypoxia-Inducible Factor-2? in von Hippel Lindau Renal Cell Carcinoma"

    Article Title: Identification of Membrane Type-1 Matrix Metalloproteinase as a Target of Hypoxia-Inducible Factor-2? in von Hippel Lindau Renal Cell Carcinoma

    Journal: Oncogene

    doi: 10.1038/sj.onc.1208305

    MT1-MMP overexpression in RCC cells lacking VHL. Total RNA and protein were isolated from WT8 and pRc-9 cells. (A) Northern blot analysis of MT1-MMP mRNA expression and western blot analysis of VHL protein expression in RCC cell lines. RNA levels were
    Figure Legend Snippet: MT1-MMP overexpression in RCC cells lacking VHL. Total RNA and protein were isolated from WT8 and pRc-9 cells. (A) Northern blot analysis of MT1-MMP mRNA expression and western blot analysis of VHL protein expression in RCC cell lines. RNA levels were

    Techniques Used: Over Expression, Isolation, Northern Blot, Expressing, Western Blot

    Related Articles

    Transduction:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α. .. The resulting pKR13 was moved into S. aureus RN4220 and UAMS-1 by electroporation and phage transduction, respectively, as described above.

    Clone Assay:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: Each PCR product was cloned into pCRBlunt (Invitrogen Life Technologies) following the manufacturer's protocols for ligation and E. coli transformation, followed by Sanger DNA sequencing (ICBR, University of Florida) to verify that no mutations had been introduced. .. Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α.

    Southern Blot:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: PCR using the nos1-F and nos1-R primer pair, as well as Southern blotting, were used to screen candidate mutants to confirm that the chromosomal nos gene had been correctly replaced by the nos :: erm allele. .. Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α.

    Ligation:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: .. Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α. .. The resulting pKR13 was moved into S. aureus RN4220 and UAMS-1 by electroporation and phage transduction, respectively, as described above.

    Mutagenesis:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: Paragraph title: Creation of nos ::erm and Δpdt mutant strains ... Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α.

    Article Title: Identification of Membrane Type-1 Matrix Metalloproteinase as a Target of Hypoxia-Inducible Factor-2? in von Hippel Lindau Renal Cell Carcinoma
    Article Snippet: The oligos used in the mutagenesis were designed from the MT1-MMP sequence (GenBank accession # ) using Stratagene’s web-based primer design software found at . .. The pTK-RE3-luc, pCMV-HIF-2α, and pCMV-HIFβ expression constructs were generous gifts of Richard Bruick (University of Texas, Southwestern Medical Center, Dallas, TX). pRc/CMV-HA-VHL expression construct was a kind gift from William Kaelin. pRc/CMV empty vector was created from pRc/CMV-HA-VHL by HindIII/XbaI digest to remove the HA-VHL insert and was religated using HindIII-XmnI + XbaI-XmnI adaptors (NE Biolabs).

    Isolation:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: On days 3–5, the culture was serially-diluted and spread on TSA +10 µg/ml Erm, and isolated colonies were then screened for Erm-resistant and Cm-sensitive phenotypes by picking and patching onto TSA +2 µg/ml Erm and TSA +5 µg/ml Cm. .. Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α.

    Construct:

    Article Title: Identification of Membrane Type-1 Matrix Metalloproteinase as a Target of Hypoxia-Inducible Factor-2? in von Hippel Lindau Renal Cell Carcinoma
    Article Snippet: .. The pTK-RE3-luc, pCMV-HIF-2α, and pCMV-HIFβ expression constructs were generous gifts of Richard Bruick (University of Texas, Southwestern Medical Center, Dallas, TX). pRc/CMV-HA-VHL expression construct was a kind gift from William Kaelin. pRc/CMV empty vector was created from pRc/CMV-HA-VHL by HindIII/XbaI digest to remove the HA-VHL insert and was religated using HindIII-XmnI + XbaI-XmnI adaptors (NE Biolabs). .. Total cellular RNA was purified from using TRIzol Reagent (Invitrogen).

    Polymerase Chain Reaction:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: Each PCR product was cloned into pCRBlunt (Invitrogen Life Technologies) following the manufacturer's protocols for ligation and E. coli transformation, followed by Sanger DNA sequencing (ICBR, University of Florida) to verify that no mutations had been introduced. .. Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α.

    Incubation:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: .. Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α. .. The resulting pKR13 was moved into S. aureus RN4220 and UAMS-1 by electroporation and phage transduction, respectively, as described above.

    DNA Sequencing:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: Each PCR product was cloned into pCRBlunt (Invitrogen Life Technologies) following the manufacturer's protocols for ligation and E. coli transformation, followed by Sanger DNA sequencing (ICBR, University of Florida) to verify that no mutations had been introduced. .. Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α.

    Expressing:

    Article Title: Identification of Membrane Type-1 Matrix Metalloproteinase as a Target of Hypoxia-Inducible Factor-2? in von Hippel Lindau Renal Cell Carcinoma
    Article Snippet: .. The pTK-RE3-luc, pCMV-HIF-2α, and pCMV-HIFβ expression constructs were generous gifts of Richard Bruick (University of Texas, Southwestern Medical Center, Dallas, TX). pRc/CMV-HA-VHL expression construct was a kind gift from William Kaelin. pRc/CMV empty vector was created from pRc/CMV-HA-VHL by HindIII/XbaI digest to remove the HA-VHL insert and was religated using HindIII-XmnI + XbaI-XmnI adaptors (NE Biolabs). .. Total cellular RNA was purified from using TRIzol Reagent (Invitrogen).

    Gel Extraction:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: The pdt1 and pdt2 inserts were each excised from pCRBlunt by restriction digestion with HindIII-ClaI and ClaI-XbaI, respectively, followed by gel-purification of each insert (DNA Gel Extraction Kit, Zymo Research). .. Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α.

    Sequencing:

    Article Title: Identification of Membrane Type-1 Matrix Metalloproteinase as a Target of Hypoxia-Inducible Factor-2? in von Hippel Lindau Renal Cell Carcinoma
    Article Snippet: Mutations were confirmed by DNA sequence analysis. pTK-RE3-luc reporter plasmid contains 3 copies of a 50nt hypoxia-inducible enhancer from the erythropoietin gene and has been shown to be inducible by co-expression of HIF-2α ( ). .. The pTK-RE3-luc, pCMV-HIF-2α, and pCMV-HIFβ expression constructs were generous gifts of Richard Bruick (University of Texas, Southwestern Medical Center, Dallas, TX). pRc/CMV-HA-VHL expression construct was a kind gift from William Kaelin. pRc/CMV empty vector was created from pRc/CMV-HA-VHL by HindIII/XbaI digest to remove the HA-VHL insert and was religated using HindIII-XmnI + XbaI-XmnI adaptors (NE Biolabs).

    Electroporation:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α. .. The resulting pKR13 was moved into S. aureus RN4220 and UAMS-1 by electroporation and phage transduction, respectively, as described above.

    Transformation Assay:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: .. Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α. .. The resulting pKR13 was moved into S. aureus RN4220 and UAMS-1 by electroporation and phage transduction, respectively, as described above.

    Gel Purification:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: The pdt1 and pdt2 inserts were each excised from pCRBlunt by restriction digestion with HindIII-ClaI and ClaI-XbaI, respectively, followed by gel-purification of each insert (DNA Gel Extraction Kit, Zymo Research). .. Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α.

    Plasmid Preparation:

    Article Title: Contribution of the nos-pdt Operon to Virulence Phenotypes in Methicillin-Sensitive Staphylococcus aureus
    Article Snippet: .. Plasmid pBT2 was digested with HindIII-XbaI, and a “triple” ligation reaction using T4 DNA ligase (New England Biolabs) containing this linearized plasmid, HindIII-ClaI pdt1 and ClaI-XbaI pdt2 was incubated for several hours at room temperature, followed by heat-shock transformation into E. coli DH5α. .. The resulting pKR13 was moved into S. aureus RN4220 and UAMS-1 by electroporation and phage transduction, respectively, as described above.

    Article Title: Identification of Membrane Type-1 Matrix Metalloproteinase as a Target of Hypoxia-Inducible Factor-2? in von Hippel Lindau Renal Cell Carcinoma
    Article Snippet: .. The pTK-RE3-luc, pCMV-HIF-2α, and pCMV-HIFβ expression constructs were generous gifts of Richard Bruick (University of Texas, Southwestern Medical Center, Dallas, TX). pRc/CMV-HA-VHL expression construct was a kind gift from William Kaelin. pRc/CMV empty vector was created from pRc/CMV-HA-VHL by HindIII/XbaI digest to remove the HA-VHL insert and was religated using HindIII-XmnI + XbaI-XmnI adaptors (NE Biolabs). .. Total cellular RNA was purified from using TRIzol Reagent (Invitrogen).

    Software:

    Article Title: Identification of Membrane Type-1 Matrix Metalloproteinase as a Target of Hypoxia-Inducible Factor-2? in von Hippel Lindau Renal Cell Carcinoma
    Article Snippet: The oligos used in the mutagenesis were designed from the MT1-MMP sequence (GenBank accession # ) using Stratagene’s web-based primer design software found at . .. The pTK-RE3-luc, pCMV-HIF-2α, and pCMV-HIFβ expression constructs were generous gifts of Richard Bruick (University of Texas, Southwestern Medical Center, Dallas, TX). pRc/CMV-HA-VHL expression construct was a kind gift from William Kaelin. pRc/CMV empty vector was created from pRc/CMV-HA-VHL by HindIII/XbaI digest to remove the HA-VHL insert and was religated using HindIII-XmnI + XbaI-XmnI adaptors (NE Biolabs).

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    New England Biolabs hindiii xmni xbai xmni adaptors
    Hindiii Xmni Xbai Xmni Adaptors, supplied by New England Biolabs, 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/hindiii xmni xbai xmni adaptors/product/New England Biolabs
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hindiii xmni xbai xmni adaptors - by Bioz Stars, 2020-04
    85/100 stars
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