摘要
目的构建乙型脑炎病毒NS1基因的重组原核表达质粒,诱导重组蛋白表达及免疫学鉴定。方法以乙型脑炎病毒基因组为模板,RT-PCR扩增NS1基因片段,构建其重组原核表达载体,并采用WesternBlot方法初步鉴定该蛋白的特异性。结果成功扩增出乙型脑炎病毒NS1基因片段,该基因在大肠杆菌表达系统中得到表达,并且特异性识别乙型脑炎患者血清。结论在大肠杆菌中成功表达乙型脑炎病毒NS1基因片段,为乙型脑炎病毒的诊断提供了新的特异性抗原。
Objective To clone, express and identify the NS1 gene from Japanese encephalitis virus. Methods NSI gene was amplified by the RT-PCR reaction from Japanese encephalitis virus and cloned into pGEX-4T-2 vector. After induction by IPTG, the NS1 protein was identified by Western Blot. Results The NS1 gene （420 bp） was amplified and the recombinant protein was expressed successfully. The result of Western Blot showed that recombinant NS 1 protein was identified specifically by serum of Japanese encephalitis virus. Conclusion The recombinant NS 1 protein was expressed successfully and contributed to the diagnosis of Japanese encephalitis virus.
出处
《分子诊断与治疗杂志》
2011年第6期381-383,共3页
Journal of Molecular Diagnostics and Therapy
基金
国家科技重大专项(2009ZX10004-706)
关键词
乙型脑炎病毒
NS1基因
蛋白表达
Japanese encephalitis virus
NS 1 gene
Protein expression
