蓝舌病1型病毒VP2蛋白的原核表达及免疫反应性分析

doi:10.16431/j.cnki.1671-7236.2017.07.030

Abstract

Abstract:

The study was aimed to test the immunoreactivity of the VP2 protein of bluetongue virus serotype 1 (BTV-1) in vitro. Based on the published BTV-1 L2 gene of Y863 strain, specific cloning PCR primers were designed and synthesized. The L2 gene was amplified through RT-PCR method and then was purified and cloned into the expressing vector pEASY-Blunt E1. The cloned recombinant plasmids were identified. The positive recombinant L2 plasmid was cloned into BL21(DE3) competent cells to express VP2 protein. The acquired purified recombinant BTV-1 VP2 protein was analyzed through the methods of Western blotting, ELISA and blocking ELISA. The results showed that:BTV-1 VP2 protein was expressed as the inclusion bodies in the pEASY-Blunt E1 vector; 160 and 200 mmol/L glyoxaline were the best condition to wash down the expressed protein. The molecular weight of this purified recombinant protein with N-terminal His-tag was about 105 ku. Through the results of Western blotting, ELISA and blocking ELISA, it had been proved that this recombinant protein could combine with BTV-1 specific antibody and this combination could be blocked by BTV-1 virus. The study showed that the recombinant BTV-1 VP2 protein, expressed through the prokaryotic expression vector pEASY-Blunt E1, possessed good immunoreactivity and this study had established foundation for locating the serotypic epitopes of the BTV-1 VP2 protein.

Key words: bluetongue; VP2 protein; expression; immunoreactivity

CLC Number:

S852.65

SONG Jian-ling, LI Hua-chun. Prokaryotic Expression and Immunoreactivity Analysis of VP2 Protein of Bluetongue Virus Serotype 1[J]. , 2017, 44(7): 2119-2125.

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Prokaryotic Expression and Immunoreactivity Analysis of VP2 Protein of Bluetongue Virus Serotype 1

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