Prokaryotic Expression of the GP3 Protein of Porcine Reproductive and Respiratory Syndrome Virus S1 Strain

LI Pei; CAO Rui-bing; ZHOU Bin; ZHENG Qi-sheng; WEI Jian-chao; LI Peng; REN Xue-feng; CHEN Fu-yan

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October 2005

LI Pei, CAO Rui-bing, ZHOU Bin, ZHENG Qi-sheng, WEI Jian-chao, LI Peng, REN Xue-feng and CHEN Fu-yan. Prokaryotic Expression of the GP3 Protein of Porcine Reproductive and Respiratory Syndrome Virus S1 Strain[J]. Virologica Sinica, 2005, 20(5): 507-510.

Citation: LI Pei, CAO Rui-bing, ZHOU Bin, ZHENG Qi-sheng, WEI Jian-chao, LI Peng, REN Xue-feng, CHEN Fu-yan. Prokaryotic Expression of the GP3 Protein of Porcine Reproductive and Respiratory Syndrome Virus S1 Strain .VIROLOGICA SINICA, 2005, 20(5) : 507-510.

猪细小病毒VP2蛋白的主要抗原表位基因的原核表达及检测应用

李斐 ¹ ,
曹瑞兵 ¹ ,
周斌 ¹ ,
郑其升 ¹ ,
魏建超 ¹ ,
李鹏 ¹ ,
任雪枫 ² ,
陈溥言 ^1*

1.
1.江苏南京南京农业大学农业部畜禽疫病诊断与免疫重点开放实验室 2. 江苏省畜牧兽医总站

摘要

将构建好的重组质粒PET-VP2Ⅰ转化宿主菌BL21,利用IPTG(1mmol/L)诱导实现了PET-VP2Ⅰ蛋白主要抗原域VP2Ⅰ融合蛋白的高效表达。通过Western-blot检测证明表达的重组蛋白具有良好的免疫学活性。表达产物经HisBind层析柱纯化后作为诊断抗原初步建立了检测PPV抗体的间接ELISA方法。结果表明:抗原的最佳包被浓度为3.5μg/mL,血清的最佳稀释度为1∶40,待检血清阳性标准初步定为OD4900.51,且待检血清的OD490值与阴性血清的OD490值的比值大于2.1。
- 猪细小病毒
- , VP2蛋白的主要抗原域
- , 原核表达
- , 间接ELISA

Prokaryotic Expression of the GP3 Protein of Porcine Reproductive and Respiratory Syndrome Virus S1 Strain

Abstract

The recombinant plasmid pET-VP_2Ⅰ was transformed into BL21 competent cells and expressed in high level after induced with 1.0mmol/l IPTG.The expressed product was purified with His·Bind chromatograghy after being proved by Westen-blot,and was used as an antigen to establish indirect ELISA for detecting antibodies against PPV.The optional working circumstances for the ELISA(antigen concentration 3.5μg/mL;serum dilution 1∶40)were tried out with chess titration.The positive critertion of test serum of this ELISA is OD_490nm0.51,the OD_490 ratio of the tested serum to the negative serum is higher than 2.1.
- Porcine parvovirus（PPV）
- , Main antigenic region of VP_2 protein
- , Prokaryotic expression
- , Indirect ELISA

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Prokaryotic Expression of the GP3 Protein of Porcine Reproductive and Respiratory Syndrome Virus S1 Strain

Abstract: The recombinant plasmid pET-VP_2Ⅰ was transformed into BL21 competent cells and expressed in high level after induced with 1.0mmol/l IPTG.The expressed product was purified with His·Bind chromatograghy after being proved by Westen-blot,and was used as an antigen to establish indirect ELISA for detecting antibodies against PPV.The optional working circumstances for the ELISA(antigen concentration 3.5μg/mL;serum dilution 1∶40)were tried out with chess titration.The positive critertion of test serum of this ELISA is OD_490nm0.51,the OD_490 ratio of the tested serum to the negative serum is higher than 2.1.