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5 PRIME gpt gene
Construction of a vaccinia virus based <t>SARS-CoV</t> reverse genetic system. The genome structure of SARS-CoV is shown at the top of the figure. Nine cDNA clones produced from the genomic RNA of SARS-CoV isolate HKU-39849 are shown below. The region of the SARS-CoV genome encompassed by each clone is indicated by the nucleotide number (using the recSARS-CoV sequence; GenBank: JN854286) at the beginning and end of each clone. Restriction enzyme sites used to join the clones are shown, with restriction enzymes sites added to the clones shown in bold. The cDNA fragments isolated from the clones and <t>gpt</t> PCR products covering regions of the genome unstable as cDNA clones were ligated with each other and vaccinia virus DNA to produce two vaccinia virus recombinant clones spanning nts 1–20288 and 20272–29727 of the SARS-CoV genome respectively. The first 2012 nts of the former vaccinia virus recombinant was derived from the SARS-CoV isolate Frankfurt-1 (shaded in dark grey). Vaccinia virus mediated homologous recombination was then used to reconstitute the SARS-CoV subgenomic fragments, introducing regions of cDNA that were unstable in E. coli and repairing errors (*) introduced during the cloning process. This resulted in the vaccinia virus clones vSARS-CoV-5prime and vSARS-CoV-3prime. The SARS-CoV cDNA fragments were isolated from the two vaccinia virus recombinants by restriction enzyme digestion and then joined using unique SfiI and BglI sites that had been introduced into the cDNA. The ligated cDNA fragments were used as a template for in vitro transcription using a T7 polymerase promoter introduced at the 5′ end of the SARS-CoV 5′ cDNA clone to produce a RNA transcript representing the SARS-CoV genome.
Gpt Gene, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gpt gene/product/5 PRIME
Average 88 stars, based on 2 article reviews
Price from $9.99 to $1999.99
gpt gene - by Bioz Stars, 2020-08
88/100 stars

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1) Product Images from "Reverse Genetics of SARS-Related Coronavirus Using Vaccinia Virus-Based Recombination"

Article Title: Reverse Genetics of SARS-Related Coronavirus Using Vaccinia Virus-Based Recombination

Journal: PLoS ONE

doi: 10.1371/journal.pone.0032857

Construction of a vaccinia virus based SARS-CoV reverse genetic system. The genome structure of SARS-CoV is shown at the top of the figure. Nine cDNA clones produced from the genomic RNA of SARS-CoV isolate HKU-39849 are shown below. The region of the SARS-CoV genome encompassed by each clone is indicated by the nucleotide number (using the recSARS-CoV sequence; GenBank: JN854286) at the beginning and end of each clone. Restriction enzyme sites used to join the clones are shown, with restriction enzymes sites added to the clones shown in bold. The cDNA fragments isolated from the clones and gpt PCR products covering regions of the genome unstable as cDNA clones were ligated with each other and vaccinia virus DNA to produce two vaccinia virus recombinant clones spanning nts 1–20288 and 20272–29727 of the SARS-CoV genome respectively. The first 2012 nts of the former vaccinia virus recombinant was derived from the SARS-CoV isolate Frankfurt-1 (shaded in dark grey). Vaccinia virus mediated homologous recombination was then used to reconstitute the SARS-CoV subgenomic fragments, introducing regions of cDNA that were unstable in E. coli and repairing errors (*) introduced during the cloning process. This resulted in the vaccinia virus clones vSARS-CoV-5prime and vSARS-CoV-3prime. The SARS-CoV cDNA fragments were isolated from the two vaccinia virus recombinants by restriction enzyme digestion and then joined using unique SfiI and BglI sites that had been introduced into the cDNA. The ligated cDNA fragments were used as a template for in vitro transcription using a T7 polymerase promoter introduced at the 5′ end of the SARS-CoV 5′ cDNA clone to produce a RNA transcript representing the SARS-CoV genome.
Figure Legend Snippet: Construction of a vaccinia virus based SARS-CoV reverse genetic system. The genome structure of SARS-CoV is shown at the top of the figure. Nine cDNA clones produced from the genomic RNA of SARS-CoV isolate HKU-39849 are shown below. The region of the SARS-CoV genome encompassed by each clone is indicated by the nucleotide number (using the recSARS-CoV sequence; GenBank: JN854286) at the beginning and end of each clone. Restriction enzyme sites used to join the clones are shown, with restriction enzymes sites added to the clones shown in bold. The cDNA fragments isolated from the clones and gpt PCR products covering regions of the genome unstable as cDNA clones were ligated with each other and vaccinia virus DNA to produce two vaccinia virus recombinant clones spanning nts 1–20288 and 20272–29727 of the SARS-CoV genome respectively. The first 2012 nts of the former vaccinia virus recombinant was derived from the SARS-CoV isolate Frankfurt-1 (shaded in dark grey). Vaccinia virus mediated homologous recombination was then used to reconstitute the SARS-CoV subgenomic fragments, introducing regions of cDNA that were unstable in E. coli and repairing errors (*) introduced during the cloning process. This resulted in the vaccinia virus clones vSARS-CoV-5prime and vSARS-CoV-3prime. The SARS-CoV cDNA fragments were isolated from the two vaccinia virus recombinants by restriction enzyme digestion and then joined using unique SfiI and BglI sites that had been introduced into the cDNA. The ligated cDNA fragments were used as a template for in vitro transcription using a T7 polymerase promoter introduced at the 5′ end of the SARS-CoV 5′ cDNA clone to produce a RNA transcript representing the SARS-CoV genome.

Techniques Used: Clone Assay, Produced, Sequencing, Isolation, Polymerase Chain Reaction, Recombinant, Derivative Assay, Homologous Recombination, In Vitro

2) Product Images from "Reverse Genetics of SARS-Related Coronavirus Using Vaccinia Virus-Based Recombination"

Article Title: Reverse Genetics of SARS-Related Coronavirus Using Vaccinia Virus-Based Recombination

Journal: PLoS ONE

doi: 10.1371/journal.pone.0032857

Construction of a vaccinia virus based SARS-CoV reverse genetic system. The genome structure of SARS-CoV is shown at the top of the figure. Nine cDNA clones produced from the genomic RNA of SARS-CoV isolate HKU-39849 are shown below. The region of the SARS-CoV genome encompassed by each clone is indicated by the nucleotide number (using the recSARS-CoV sequence; GenBank: JN854286) at the beginning and end of each clone. Restriction enzyme sites used to join the clones are shown, with restriction enzymes sites added to the clones shown in bold. The cDNA fragments isolated from the clones and gpt PCR products covering regions of the genome unstable as cDNA clones were ligated with each other and vaccinia virus DNA to produce two vaccinia virus recombinant clones spanning nts 1–20288 and 20272–29727 of the SARS-CoV genome respectively. The first 2012 nts of the former vaccinia virus recombinant was derived from the SARS-CoV isolate Frankfurt-1 (shaded in dark grey). Vaccinia virus mediated homologous recombination was then used to reconstitute the SARS-CoV subgenomic fragments, introducing regions of cDNA that were unstable in E. coli and repairing errors (*) introduced during the cloning process. This resulted in the vaccinia virus clones vSARS-CoV-5prime and vSARS-CoV-3prime. The SARS-CoV cDNA fragments were isolated from the two vaccinia virus recombinants by restriction enzyme digestion and then joined using unique SfiI and BglI sites that had been introduced into the cDNA. The ligated cDNA fragments were used as a template for in vitro transcription using a T7 polymerase promoter introduced at the 5′ end of the SARS-CoV 5′ cDNA clone to produce a RNA transcript representing the SARS-CoV genome.
Figure Legend Snippet: Construction of a vaccinia virus based SARS-CoV reverse genetic system. The genome structure of SARS-CoV is shown at the top of the figure. Nine cDNA clones produced from the genomic RNA of SARS-CoV isolate HKU-39849 are shown below. The region of the SARS-CoV genome encompassed by each clone is indicated by the nucleotide number (using the recSARS-CoV sequence; GenBank: JN854286) at the beginning and end of each clone. Restriction enzyme sites used to join the clones are shown, with restriction enzymes sites added to the clones shown in bold. The cDNA fragments isolated from the clones and gpt PCR products covering regions of the genome unstable as cDNA clones were ligated with each other and vaccinia virus DNA to produce two vaccinia virus recombinant clones spanning nts 1–20288 and 20272–29727 of the SARS-CoV genome respectively. The first 2012 nts of the former vaccinia virus recombinant was derived from the SARS-CoV isolate Frankfurt-1 (shaded in dark grey). Vaccinia virus mediated homologous recombination was then used to reconstitute the SARS-CoV subgenomic fragments, introducing regions of cDNA that were unstable in E. coli and repairing errors (*) introduced during the cloning process. This resulted in the vaccinia virus clones vSARS-CoV-5prime and vSARS-CoV-3prime. The SARS-CoV cDNA fragments were isolated from the two vaccinia virus recombinants by restriction enzyme digestion and then joined using unique SfiI and BglI sites that had been introduced into the cDNA. The ligated cDNA fragments were used as a template for in vitro transcription using a T7 polymerase promoter introduced at the 5′ end of the SARS-CoV 5′ cDNA clone to produce a RNA transcript representing the SARS-CoV genome.

Techniques Used: Clone Assay, Produced, Sequencing, Isolation, Polymerase Chain Reaction, Recombinant, Derivative Assay, Homologous Recombination, In Vitro

Related Articles

Recombinant:

Article Title: Reverse Genetics of SARS-Related Coronavirus Using Vaccinia Virus-Based Recombination
Article Snippet: .. Thus, initially, we substituted this region of the SARS-CoV cDNA with a gpt gene in the recombinant vaccinia virus, vSARS-CoV-5prime-gpt. .. Subsequently, the gpt gene was replaced by the appropriate SARS-CoV cDNA sequences using homologous recombination involving vaccinia virus vSARS-CoV-5prime-gpt and a plasmid DNA that contained a shorter region of SARS-CoV cDNA (nts 10808 to 12837).

Homologous Recombination:

Article Title: Reverse Genetics of SARS-Related Coronavirus Using Vaccinia Virus-Based Recombination
Article Snippet: .. Subsequently, the gpt gene was replaced by the appropriate SARS-CoV cDNA sequences using homologous recombination involving vaccinia virus vSARS-CoV-5prime-gpt and a plasmid DNA that contained a shorter region of SARS-CoV cDNA (nts 10808 to 12837). .. Homologous recombination was also used to repair a non-silent nucleotide change corresponding to SARS-CoV nt 18292 in vSARS-CoV-5prime.

Plasmid Preparation:

Article Title: Reverse Genetics of SARS-Related Coronavirus Using Vaccinia Virus-Based Recombination
Article Snippet: .. Subsequently, the gpt gene was replaced by the appropriate SARS-CoV cDNA sequences using homologous recombination involving vaccinia virus vSARS-CoV-5prime-gpt and a plasmid DNA that contained a shorter region of SARS-CoV cDNA (nts 10808 to 12837). .. Homologous recombination was also used to repair a non-silent nucleotide change corresponding to SARS-CoV nt 18292 in vSARS-CoV-5prime.

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    5 PRIME gpt gene
    Construction of a vaccinia virus based <t>SARS-CoV</t> reverse genetic system. The genome structure of SARS-CoV is shown at the top of the figure. Nine cDNA clones produced from the genomic RNA of SARS-CoV isolate HKU-39849 are shown below. The region of the SARS-CoV genome encompassed by each clone is indicated by the nucleotide number (using the recSARS-CoV sequence; GenBank: JN854286) at the beginning and end of each clone. Restriction enzyme sites used to join the clones are shown, with restriction enzymes sites added to the clones shown in bold. The cDNA fragments isolated from the clones and <t>gpt</t> PCR products covering regions of the genome unstable as cDNA clones were ligated with each other and vaccinia virus DNA to produce two vaccinia virus recombinant clones spanning nts 1–20288 and 20272–29727 of the SARS-CoV genome respectively. The first 2012 nts of the former vaccinia virus recombinant was derived from the SARS-CoV isolate Frankfurt-1 (shaded in dark grey). Vaccinia virus mediated homologous recombination was then used to reconstitute the SARS-CoV subgenomic fragments, introducing regions of cDNA that were unstable in E. coli and repairing errors (*) introduced during the cloning process. This resulted in the vaccinia virus clones vSARS-CoV-5prime and vSARS-CoV-3prime. The SARS-CoV cDNA fragments were isolated from the two vaccinia virus recombinants by restriction enzyme digestion and then joined using unique SfiI and BglI sites that had been introduced into the cDNA. The ligated cDNA fragments were used as a template for in vitro transcription using a T7 polymerase promoter introduced at the 5′ end of the SARS-CoV 5′ cDNA clone to produce a RNA transcript representing the SARS-CoV genome.
    Gpt Gene, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gpt gene/product/5 PRIME
    Average 88 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    gpt gene - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

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    Construction of a vaccinia virus based SARS-CoV reverse genetic system. The genome structure of SARS-CoV is shown at the top of the figure. Nine cDNA clones produced from the genomic RNA of SARS-CoV isolate HKU-39849 are shown below. The region of the SARS-CoV genome encompassed by each clone is indicated by the nucleotide number (using the recSARS-CoV sequence; GenBank: JN854286) at the beginning and end of each clone. Restriction enzyme sites used to join the clones are shown, with restriction enzymes sites added to the clones shown in bold. The cDNA fragments isolated from the clones and gpt PCR products covering regions of the genome unstable as cDNA clones were ligated with each other and vaccinia virus DNA to produce two vaccinia virus recombinant clones spanning nts 1–20288 and 20272–29727 of the SARS-CoV genome respectively. The first 2012 nts of the former vaccinia virus recombinant was derived from the SARS-CoV isolate Frankfurt-1 (shaded in dark grey). Vaccinia virus mediated homologous recombination was then used to reconstitute the SARS-CoV subgenomic fragments, introducing regions of cDNA that were unstable in E. coli and repairing errors (*) introduced during the cloning process. This resulted in the vaccinia virus clones vSARS-CoV-5prime and vSARS-CoV-3prime. The SARS-CoV cDNA fragments were isolated from the two vaccinia virus recombinants by restriction enzyme digestion and then joined using unique SfiI and BglI sites that had been introduced into the cDNA. The ligated cDNA fragments were used as a template for in vitro transcription using a T7 polymerase promoter introduced at the 5′ end of the SARS-CoV 5′ cDNA clone to produce a RNA transcript representing the SARS-CoV genome.

    Journal: PLoS ONE

    Article Title: Reverse Genetics of SARS-Related Coronavirus Using Vaccinia Virus-Based Recombination

    doi: 10.1371/journal.pone.0032857

    Figure Lengend Snippet: Construction of a vaccinia virus based SARS-CoV reverse genetic system. The genome structure of SARS-CoV is shown at the top of the figure. Nine cDNA clones produced from the genomic RNA of SARS-CoV isolate HKU-39849 are shown below. The region of the SARS-CoV genome encompassed by each clone is indicated by the nucleotide number (using the recSARS-CoV sequence; GenBank: JN854286) at the beginning and end of each clone. Restriction enzyme sites used to join the clones are shown, with restriction enzymes sites added to the clones shown in bold. The cDNA fragments isolated from the clones and gpt PCR products covering regions of the genome unstable as cDNA clones were ligated with each other and vaccinia virus DNA to produce two vaccinia virus recombinant clones spanning nts 1–20288 and 20272–29727 of the SARS-CoV genome respectively. The first 2012 nts of the former vaccinia virus recombinant was derived from the SARS-CoV isolate Frankfurt-1 (shaded in dark grey). Vaccinia virus mediated homologous recombination was then used to reconstitute the SARS-CoV subgenomic fragments, introducing regions of cDNA that were unstable in E. coli and repairing errors (*) introduced during the cloning process. This resulted in the vaccinia virus clones vSARS-CoV-5prime and vSARS-CoV-3prime. The SARS-CoV cDNA fragments were isolated from the two vaccinia virus recombinants by restriction enzyme digestion and then joined using unique SfiI and BglI sites that had been introduced into the cDNA. The ligated cDNA fragments were used as a template for in vitro transcription using a T7 polymerase promoter introduced at the 5′ end of the SARS-CoV 5′ cDNA clone to produce a RNA transcript representing the SARS-CoV genome.

    Article Snippet: Subsequently, the gpt gene was replaced by the appropriate SARS-CoV cDNA sequences using homologous recombination involving vaccinia virus vSARS-CoV-5prime-gpt and a plasmid DNA that contained a shorter region of SARS-CoV cDNA (nts 10808 to 12837).

    Techniques: Clone Assay, Produced, Sequencing, Isolation, Polymerase Chain Reaction, Recombinant, Derivative Assay, Homologous Recombination, In Vitro