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- W213573690 abstract "Objective To clone the selected gene of the single-chain fragment of the variable region(scFv) against the nucleocapsid protein(NP) antigen of hantavirus into the prokaryotic expression vector pGEX-6p-1 to construct a recombinant expression plasmid pGEX-6P-1-scFv.Methods T7 phage DNA containing the gene for the scFv against the hantavirus NP antigen was extracted.The scFv gene was amplified with template to T7 DNA by PCR and was then digested with BamHⅠ and SalⅠ.The obtained fragment was then ligated into the expression vector pGEX-6p-1 that had been digested with BamHⅠ and SalⅠ.After the recombinant DNA was transformed into competent E.coli BL-21(DE3),a positive recombinant plasmid was selected preliminarily by agarose gel electrophoresis.The recombinant plasmid was identified by PCR,digestion of BamHⅠand SalⅠand DNA sequencing.Results The recombinant pGEX-6P-1-scFv was digested into two fragments of 5 000 bp and 750 bp by BamHⅠand SalⅠ as we had expected.The PCR product of the recombinant plasmid was 750 bp as we had expected.The scFv gene in the recombinant plasmid was sequenced and compared with the scFv gene in T7 phage DNA and was found to corresponded with the later.The results show that the gene for the scFv against NP was cloned into the expression vector pGEX-6p-1.Conclusion Prokaryotic expression recombinant pGEX-6P-1-scFv,which contained the gene for the scFv against the hantavirus NP antigen was constructed successfully." @default.
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- W213573690 date "2009-01-01" @default.
- W213573690 modified "2023-09-24" @default.
- W213573690 title "Gene cloning and identification of scFv against nucleocapsid protein of hantavirus." @default.
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