肾综合征出血热病毒核蛋白在大肠杆菌中表达及其产物的初步应用  

Expression of the nucleocapsid protein of Hantaan Virus in E. coli and preliminary use of the ex-pressed protein in diagnostic ELISA

作　　者：黄长形[1] 李盈盈[1] 杨为松[1] 杭长寿[1] 白雪帆[1] 尚高峰[1] 张文彬[1] 

机构地区：[1]西安第四军医大学唐都医院,中国预防医学科学院病毒研究所

出　　处：《中华传染病杂志》1996年第1期28-31,共4页Chinese Journal of Infectious Diseases

摘　　要：为了高效表达肾综合征出血热病毒核蛋白，并将其应用于该病的血清学诊断中，利用聚合酶链反应（ＰＣＲ）扩增了７６－ｌｌ８株病毒核蛋白编码基因并除去了非编码区基因，将编码基因正确插入表达载体ＰＢＶ２２０，经ＥＬＩＳＡ检测和蛋白免疫印迹分析，结果核蛋白得到了有效表达，表达的核蛋白占菌体总蛋白的７．５％。利用尿素变性的表达核蛋白做包被抗原，建立了检测患者血清中抗ＨＦＲＳＶ核蛋白抗体ＩｇＧ和ＩｇＭ的间接ＥＬＩＳＡ方法，提示该方法特异、稳定且敏感性较好，其优点在于简便、快速，适合于基层应用。In order to obtain high expression of nucleoprotein(NP)of Hantaan virus and to use expressed protein in diagnostic ELISA,the coding frame of NP of Hantaan virus was amplified from the cDNA of S genome segment.The NP expression plasmid was constructed by cloning the product to EcoRI and SaII site of the expressive vector PBV220 ,and the expression of NP was identified in E.coli by ELISA and Western-blot.The expression level in DH5a was about 7.5%.he expressed NP was divided into soluble and insoluble fractions,and the insoluble NP could dissolve in 4 mo1/L urea solution.An indirect ELISA was developed by using the expressed NP and was denatured in 4 mol/L urea solution as coating antigen to detect the antibody lgG and lgM against HFRSV in patient serum.The specificity and stability of the test was good,and might serve as a potential rapid early diagnostic test of HFRS.

关 键 词：肾综合征出血热 病毒 核蛋白 大肠杆菌 ELISA 

分 类 号：R512.806[医药卫生—内科学] R692[医药卫生—临床医学]