牛流行热病毒糖蛋白基因的原核表达与抗原性鉴定

doi:10.16431/j.cnki.1671-7236.2015.09.005

Abstract

Abstract: In order to explore specific candidate gene of antigens and diagnostics development for the bovine ephemeral fever virus (BEFV), the target gene sequences were derived from G gene by PCR amplification using two specific primers.The PCR products were digested by Xho Ⅰ and Nde Ⅰ and cloned into pET-30 vector, and the recombinant plasmids (480-pET-30, 1107-pET-30) were transformed into E.coil BL21 (DE3) cells, the D_{600 nm} of positive strains were 0.6 to 1.0.The recombinant strains were induced by IPTG (1.0 mmol/L) at 37 ℃.The characteristics of the target proteins were analyzed using SDS-PAGE and the immunogenicity was analyzed through Western blotting.At the same time, the target proteins were used as coating antigens to do ELISA and all rabbits were inoculated with recombinant proteins.The results showed that the expression feature of 1 107 bp gene fragment of G gene for the first time was segmented in vitro as well as has nicer biological activity and specificity, and it more be suitable as a candidate gene for molecular vaccine and diagnostics development.This study provided the theoretical foundation of the establishment of diagnostics technique and vaccine for BEFV.

Key words: bovine ephemeral fever virus (BEFV); prokaryotic expression; immunogenicity analysis

CLC Number:

S852.65

LI Zhi, ZHENG Fu-ying, GAO Shan-dian, WANG Su-yan, YUE Cheng, YIN Hong. Prokaryotic Expression and Antigenic Characterization Identification of G Gene for Bovine Ephemeral Fever Virus[J]. , 2015, 42(9): 2246-2253.

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Prokaryotic Expression and Antigenic Characterization Identification of G Gene for Bovine Ephemeral Fever Virus

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