非洲猪瘟P72基因在Bac to Bac系统中的表达及抗原性鉴定

Abstract

Abstract: This experiment was designed to express P72 gene of African swine fever virus (ASFV) in Bac to Bac system for further research in the structure. function and immunogenicity of the P72 gene. In this study,P72 gene complete sequence (1941 bp) was amplified by PCR using pGEX-6p-1-P72 for its template. The product was cloned into pFastBac HTa granulosis virus vector and transferred into DH10Bac competent cells. Recombined plasmid transfered into the Bacmid was named rBacmid-P72 and the rBacmid-P72 was transfected into Sf9 incect cells. The recombinant baculovirus was confirmed by PCR using M13 primers. Western blotting and sandwich ELISA confirmed that the P72 protein was expressed in Sf9 cells correctly and can be recognized by rabbit anti ASFV positive serum and P72 mAb. The results indicated that P72 protein expressed in Bac to Bac system had intensive specificity,stabile activity and good antigenicity,and can be used as candidate antigen for detecting ASF.

Key words: ASFV; P72 gene; Bac to Bac; antigenicity

CLC Number:

BAI Li-hua;HOU Shao-hua;JIA Hong;YUAN Wei-feng;GUO Xiao-yu;LIANG Qian-qian;ZHU Hong-fei. Bac to Bac Expression and Antigenicity Identification of P72 Gene of ASFV[J]. China Animal Husbandry & Veterinary Medicine, 2012, 39(3): 19-23.

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Bac to Bac Expression and Antigenicity Identification of P72 Gene of ASFV

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