【摘要】 目的　获得汉坦病毒重组核蛋白 ,并将其应用于血清学诊断。方法　采用PCR方法 ,从带有HTN型Z10株病毒全长核蛋白基因的质粒上扩增读码框的前 35 4bp的核蛋白基因片段。将截短的核蛋白基因片段插入表达载体pGEX 2 0T ,得到重组质粒pGEX2 0 Z10trNP ,转化大肠杆菌BL2 1(DE3) ,经IPTG诱导表达 ,表达产物经Glu tathioneSepharose 4B柱纯化 ,并进行抗原性及抗原特异性检测。结果　pGEX2 0 Z10trBP表达产物是相对分子质量约为 4 0 0 0 0的谷光甘肽转移酶 (GST)融合蛋白GST Z10trNP。经Westernblot分析 ,GST Z10trNP具有良好的抗原性。用纯化GST Z10trNP为抗原 ,用ELISA间接法检测出血热病人血清、出血热疫苗免疫的家兔及小鼠血清 ,均能很好地区分阴性和阳性血清。结论　GST Z10trNP表达量高 ,易于纯化 ,并且具有良好的抗原性及抗原特异性 ,是一种安全、廉价的诊断抗原。更多还原

【Abstract】 Objective To obtain the recombinant nucleocapsid protein of Hantavirus and use it as an antigen for serological diagnosis.Methods A truncated nucleocapsid protein gene fragment encoding aa 1 118 was amplified from a recombinant plasmid containing the full length of nucleocapsid protein gene of Hantavirus strain Z10 by PCR,and inserted into expression vector pGEX 20T to construct a recombinant plasmid pGEX20 Z10trNP,then transformed to E.coli BL21(DE3) and expressed under induction of IPTG.The expressed product was purified by Glutathione Sepharose 4B column chromatography and detected for antigenicity and specificity.Results A recombinant protein GST Z10trNP,with a relative molecular weight of 40000,was expressed.Western blot showed good antigenicity of the expressed product.The purified GST Z10trNP was used as an antigen for detecting the sera from patients and the rabbits and mice immunized with heamorrhagic fever vaccine and showed good specificity.Conclusion The expression level of GST Z10trNP was high.The expressed product was easy to be purified and showed good antigenicity and specificity.It was a safe and cheap antigen for serological diagnosis更多还原