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Position: Home > Articles > Soluble expression and antigenic analysis of NS1 protein of Japanese encephalitis virus Heilongjiang Animal Science and Veterinary Medicine 2012 (1) 17-19+157

乙型脑炎病毒NS1蛋白的可溶性表达与抗原性分析

作  者:
刘立科;陈振师;任玉红;步志高;华荣虹
单  位:
中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室;河北农业大学动物科技学院中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室
关键词:
乙型脑炎病毒;NS1蛋白;原核表达
摘  要:
为获得可溶性表达的乙型脑炎病毒(JEV)NS1蛋白,将编码JEV SA14-14-2株NS1蛋白基因克隆至原核表达载体pMAL-c5X中,构建重组融合表达质粒pc5X-NS1;用重组质粒转化宿主菌ER2523后,经IPTG诱导得到可溶性的融合蛋白MBP-NS1;优化诱导表达条件,于28℃、0.3 mmol/L IPTG诱导4 h;最后融合蛋白经直链淀粉树脂柱纯化。结果表明:重组融合蛋白MBP-NS1具有良好的抗原性和特异性。
译  名:
Soluble expression and antigenic analysis of NS1 protein of Japanese encephalitis virus
作  者:
LIU Li-ke1,2,CHEN Zhen-shi1,2,REN Yu-hong1,BU Zhi-gao2,HUA Rong-hong2(1.College of Animal Science and Technology,Hebei Agricultural University,Baoding 071001,China; 2.State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China)
关键词:
Japanese encephalitis virus;NS1 protein;prokaryotic expression
摘  要:
To obtain the expression of NS1 protein of Japanese encephalitis virus in a soluble form,the NS1 gene of JEV strain SA14-14-2 was cloned into the expression vector pMAL-c5X,then the recombinant plasmid pc5X-NS1 was confirmed by sequencing.After the recombinant plasmid was transformed into host bacteria ER2523,the fusion protein MBP-NS1 was expressed in the soluble form and purified by induced by isopropyl beta-D-thiogalactopyranoside(IPTG),then the expression conditions were further optimized.Western-blot and indirect ELISA analysis showed that the recombinant fusion protein MBP-NS1 could be specific reactive with antibodies against JEV.This work can provide an important basis for further study on NS1 protein of Japanese encephalitis virus.

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