猪繁殖与呼吸综合征病毒NSP2抗原表位区的原核表达及免疫原性分析

doi:10.16431/j.cnki.1671-7236.2017.06.031

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (6): 1796-1803.doi: 10.16431/j.cnki.1671-7236.2017.06.031

猪繁殖与呼吸综合征病毒NSP2抗原表位区的原核表达及免疫原性分析

胡荣, 白伟杰, 王治家, 孙普, 卢曾军, 李冬, 刘在新

中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, 兰州 730046

收稿日期:2016-11-30 出版日期:2017-06-20 发布日期:2017-06-28
通讯作者: 卢曾军,李冬 E-mail:luzengjun@caas.cn;lidong@caas.cn
作者简介:胡荣(1991-),女,湖北恩施人,硕士生,研究方向:动物病毒分子生物学与免疫学,E-mail:929659925@qq.com
基金资助:
中国农业科学院创新工程

Prokaryotic Expression and Immunogenicity Analysis of Epitope Domain of NSP2 of Porcine Reproductive and Respiratory Syndrome Virus

HU Rong, BAI Wei-jie, WANG Zhi-jia, SUN Pu, LU Zeng-jun, LI Dong, LIU Zai-xin

State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

Received:2016-11-30 Online:2017-06-20 Published:2017-06-28

摘要/Abstract

摘要：

试验旨在表达纯化猪繁殖与呼吸综合征病毒NSP2重组蛋白。利用PCR技术扩增获得大小为750 bp的NSP2 226－475位氨基酸编码序列，然后将该序列亚克隆到pET-28a（+）表达载体上，通过pET-28a（+）表达载体在大肠杆菌Rosetta-gami 2（DE3）pLys中得以表达，获得大小为29 ku的重组蛋白，对重组蛋白诱导时间进行优化，SDS-PAGE显示重组蛋白在37 ℃经1 mmol/L IPTG诱导8 h时表达量最大，免疫印迹结果证实表达的蛋白能被PRRSV阳性血清识别，具有良好的免疫原性。本研究成功表达纯化了NSP2重组蛋白，为目标蛋白作为包被抗原建立检测PRRSV血清抗体的ELISA方法奠定了基础。

关键词: 猪繁殖与呼吸综合征; NSP2; 免疫原性

Abstract:

This experiment was conduced to express and purify the recombinant NSP2 protein of PRRSV. The NSP2 gene was amplified using PCR method, and then it was subcloned into pET-28a(+) vector and expressed in E.coli. The molecular weight of the recombinant protein was 29 ku. The result of SDS-PAGE indicated that the expression efficiency of the recombinant protein was induced by 1 mmol/L IPTG for 8 h at 37 ℃.Western blotting result showed that the recombinant protein had high immunogenicity and could be recognized with positive serum of PRRSV. In this study, we expressed and purified the recombinant NSP2 protein of PRRSV, and the results laid the foundation for the further development of ELISA methods coated with NSP2 protein.

Key words: porcine reproductive and respiratory syndrome; NSP2; immunogenicity

中图分类号:

S855

胡荣, 白伟杰, 王治家, 孙普, 卢曾军, 李冬, 刘在新. 猪繁殖与呼吸综合征病毒NSP2抗原表位区的原核表达及免疫原性分析[J]. 《中国畜牧兽医》, 2017, 44(6): 1796-1803.

HU Rong, BAI Wei-jie, WANG Zhi-jia, SUN Pu, LU Zeng-jun, LI Dong, LIU Zai-xin. Prokaryotic Expression and Immunogenicity Analysis of Epitope Domain of NSP2 of Porcine Reproductive and Respiratory Syndrome Virus[J]. , 2017, 44(6): 1796-1803.

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猪繁殖与呼吸综合征病毒NSP2抗原表位区的原核表达及免疫原性分析

Prokaryotic Expression and Immunogenicity Analysis of Epitope Domain of NSP2 of Porcine Reproductive and Respiratory Syndrome Virus

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