牛副流感病毒3型HNex蛋白表达及其多克隆抗体的制备

doi:10.16431/j.cnki.1671-7236.2021.01.029

Abstract

Abstract: The purpose of this study was to obtain the HNex protein of Bovine parainfluenza virus (BPIV3) and its polyclonal antibody.The extracted BPIV3 cytotoxic RNA was used as the template,and the gene fragment containing HN was amplified by RT-PCR,and then using this as the template,the HN protein outer region (HNex gene) was amplified and the nucleotide sequence was determined.The HNex gene was inserted into the cloning vector pEASY-Blunt Simple,and then double-digested into the pET-30a(+) expression vector to construct a recombinant prokaryotic expression vector pET-BPIV3-HNex and transformed into competent Rosetta^TM(DE3) pLysS.After IPTG induction,the expression products were identified by SDS-PAGE and Western blotting.HNex protein purified by affinity chromatography was used as immunogen to prepare rabbit anti-BPIV3-HNex polyclonal antibody.Sequencing results showed that this study successfully cloned the BPIV3 HNex gene.The prokaryotic expressed protein results showed that a specific band appeared at the 53 ku position,and it could specifically react with murine anti-His.The ELISA titer of BPIV3-HNex antibody in the serum of immunized rabbits was 1:819 200.The Western blotting results showed that the rabbit anti-BPIV3-HNex polyclonal antibody could specifically react with BPIV3 protein.In summary,this study used the prokaryotic expression system to express the BPIV3 HNex protein and obtained rabbit anti-BPIV3 HNex polyclonal antibodies,providing references for further exploration of BPIV3 HN protein function and the development of subunit vaccines.

Key words: Bovine parainfluenza virus type 3 (BPIV3); HNex protein; prokaryotic expression; polyclonal antibody

CLC Number:

S823

LI Mingzhu, QU Zhehui, SONG Zhifeng, LI Chenfeng, YU Yueyang, GAO Mingchun, WANG Junwei. Expression of Bovine Parainfluenza Virus Type 3 HNex Protein and Preparation of Its Polyclonal Antibodies[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 265-272.

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Expression of Bovine Parainfluenza Virus Type 3 HNex Protein and Preparation of Its Polyclonal Antibodies

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