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《Journal of Anhui Agricultural Sciences》 2008-26
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Cloning and Construction of Expression Vector of GP5 Gene of PRRSV Hn-1/06 Strain

LU Gao-feng et al(College of Animal Husbandry and Veterinary,Henan Agricultural University,Zhengzhou,Henan 450002)  
[Objective] The aim was to clone the GP5 gene of PRRSV Hn-1/06 strain and construct its prokaryotic expression vector.[Method] The fragment of GP5 gene was amplified from the Hn-1/06 strain by RT-PCR using the specific primers designed from the sequence of the ORF5 gene from PRRSV American strain ATCC-VR2332.The positive clone of PTG19-T-ORF5 was obtained after connected with PTG19-T vector.The obtained fragment of the gene was used as template to design 2 primers with the complete sequence of ORF5 and the sequence of ORF5 deleting the signal peptide resp.The target fragment was connected with the pET32a prokaryotic expression vector after PCR amplification,then identified with enzyme restriction.[Result] The GP5 gene with full length sequence of 603 bp was obtained by amplification and it encoded 200 amino acid residues and had 3 potential N linked glycosylation sites and 9 cysteines.Its molecular weight was 22.4 kD and isoelectric point was 8.550.The result of two enzyme restriction identified pET32a-ORF5 was consistent with the anticipating result,which confirmed the recombinant plasmid was constructed successfully.[Conclusion] The expression vector of GP5 gene of PRRSV Hn-1/06 strain was constructed successfully,which laid the foundation for the further study of the essence,function and the pathogenicity of GP5 protein.
【CateGory Index】: S852.65
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【Citations】
Chinese Journal Full-text Database 2 Hits
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2 Guo Baoqing(Harbin Veterinary Research Institute,the Chinese Academy of Agricultural Sciences);Isolation and Identification of Porcine Reproductory and Respiratory Syndrome(PRRS) Virus from aborted fetuses suspected of PRRS[J];;1996-02
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