V5标记的猪圆环病毒2型Cap蛋白重组杆状病毒表达载体的构建

doi:10.16431/j.cnki.1671-7236.2018.07.006

Abstract

Abstract:

The aim of the experiment was to construct the expression vector of V5-labeled porcine circovirus type 2 (PCV2) Cap protein using eukaryotic expression system.The V5 tag was introduced into the end of the PCV2 ORF2 gene using genetic engineering,and the tagged gene was cloned into the pFastBacHTA vector.Recombinant transfer vector pFastBacHTA-ORF2-V5 was transformed into E.coli DH10Bac.The identified recombinant bacmid was transfected into Sf9 cells using liposome-mediated method;Indirect immunofluorescence assay (IFA),SDS-PAGE and Western blotting were used to test the expression of V5-labeled Cap protein in Sf9 cells.The results showed that this experiment successfully introduced the V5 tag to the end of the PCV2 ORF2 gene.After the recombinant transfer vector pFastBacHTA-ORF2-V5 transformed into E.coli DH10Bac,Bacmid-ORF2-V5 recombinant bacmid was obtained.The result of IFA,SDS-PAGE and Western blotting showed that it could be expressed in Sf9 cells.The recombinant baculovirus V5-labeled PCV2 Cap protein was detected to have good reactogenicity,indicating that this experiment successfully obtained a rAcMNPV-Cap-V5 recombinant baculovirus.The test results provided a theoretical basis for the development of PCV2 marker subunit vaccine.

Key words: porcine circovirus type 2(PCV2); Cap-V5 protein; recombinant baculovirus; expression vector

CLC Number:

S852.65

XU Guo, LIANG Haiying, ZENG Zhiyong, WANG Bin, HUANG Tao, TANG Deyuan, DAI Zhenjiang, YE Baichuan, ZHANG Aiqiong, HE Xiaoli, XIAN Wen. Construction of Recombinant Baculovirus Expression Vector of V5-labeled Porcine Circovirus Type 2 Cap Protein[J]. , 2018, 45(7): 1775-1782.

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Construction of Recombinant Baculovirus Expression Vector of V5-labeled Porcine Circovirus Type 2 Cap Protein

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