动物医学进展2016，Vol.37Issue(5)：47-52,6.
猪细小病毒 VP2的原核表达与多克隆抗体制备
Prokaryotic Expression of Porcine Parvovirus VP2 and Development of Polyclonal Antibodies Against VP2
摘要
Abstract
This study aimed to clone the 1740 bp VP2 gene from porcine parvovirus,construct the prokary-otic expression vector for the expression and purification of fusion proteins and prepare polyclonal antibod-ies against porcine parvovirus VP2.Firstly,the VP2 was amplified by PCR and cloned into the prokaryotic expression vector pET-32a.Then,the recombinant vector was transformed into E.coli BL21(DE3),and the PPV VP2 fusion protein was induced by different time,IPTG concentrations,temperatures and purified by SDS-PAGE gel extraction.After three times immunization of rabbits,the antibody was prepared.The por-cine parvovirus VP2 was successfully obtained and the constructed pET-32a-VP2 was highly expressed in E.coli BL21(DE3)after induction with 1.0 mmol/L IPTG at 37℃ for 4 h.SDS-PAGE showed that the fu-sion protein was about 82 ku,and the titer of antibodies was 1:12800 by ELISA.Western blot showed that the obtained antibodies could be used to detect PPV VP2 specially and effectively.The pET-32a-VP2 was successfully constructed and VP2 fusion protein was expressed.Simultaneously,the rabbit polyclonal anti-bodies were prepared,and it laid the foundation to establish the ELISA for detecting PPV VP2 protein.关键词
猪细小病毒/VP2 蛋白/原核表达/多克隆抗体Key words
Porcine parvovirus/VP2 protein/prokaryotic expression/polyclonal antibody分类
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禹婷婷,黄勇,张樑,张洪玲,王振宇,赵志敏,童德文..猪细小病毒 VP2的原核表达与多克隆抗体制备[J].动物医学进展,2016,37(5):47-52,6.基金项目
国家自然科学基金项目 （）
