犬细小病毒VP2基因克隆及原核表达分析

doi:10.16431/j.cnki.1671-7236.2016.04.004

Abstract

Abstract: The study was aimed to investigate the epidemic situation of canine parvovirus in Hebei and Gansu provinces.VP2 protein was expressed by prokaryotic system and polyclonal antibodies were prepared, which provided a basis for the further researches on pathogenesis and therapy of canine parvovirus.Virus genomic DNA was extracted from 10 epidemic materials of suspected infected dogs and VP2 gene was amplified.The target fragments of VP2 gene were cloned to pET-30a vector after sequencing and analysis, then the positive expression plasmids were transformed into E.coli BL21 (DE3) cell.Polyclonal antibodies were prepared by immunizing guinea pigs with the SDS-PAGE identification of purified protein.The genetic analysis results showed that the VP2 gene was successfully cloned and 80% samples were belonged to CPV-2a and 20% samples were belonged to CPV-2b.The isolated strains of the test and part of China strains were gathered into a branch which had certain distance with strains of Korea and USA, and that had low homology with strains of Italy.SDS-PAGE results indicated that the recombinant protein with a molecular weight of 70 ku was expressed by prokaryotic system.Western blotting results showed that the recombinant protein had good immuneoreactivity and immunogenicity.The study indicated that CPV-2a was the predominant gene type in Hebei and Gansu provinces, meanwhile VP2 protein expressed by prokaryotic system had good antigenicity.This research provided a basis for the future study of the prevention and control of canine parvovirus.

Key words: canine parvovirus; VP2; gene type; prokaryotic expression

CLC Number:

Q785

WEI Qiao-lin, ZHANG Yun, ZHEN Shi-wei, YIN Xiang-ping. Analysis of Clone and Prokaryotic Expression of Canine Parvovirus VP2 Gene[J]. , 2016, 43(4): 870-878.

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Analysis of Clone and Prokaryotic Expression of Canine Parvovirus VP2 Gene

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