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目的:通过在原核表达系统初步构建EV71-VP1,对西安地区肠道病毒71型(EV71)的外壳蛋白VP1基因进行分析。方法:采集西安地区手足口病患儿的疱液、咽部分泌物进行病毒分离和RT-PCR检测;通过逆转录聚合酶链式反应(RT-PCR)扩增肠道病毒71型(EV71)外壳蛋白VP1基因,构建重组表达质粒PQE30/VP1,转化到大肠杆菌BL21中,对VP1基因进行遗传学分析。结果:对肠道病毒71型(EV71)西安地方株VP1基因测序,并将其与阜阳株(序列号为EU913471)相比较,表明我国西安地区EV71分离株与阜阳株有较大差别,核苷酸差异约为4%。结论:西安地方株VP1基因与阜阳株相比,其核苷酸差异较为明显。这将为西安地区EV71的分子流行病学研究以及EV71所致疾病的预防和控制,打下良好的基础。
OBJECTIVE: To analyze the coat protein VP1 gene of enterovirus 71 (EV71) in Xi’an by constructing EV71-VP1 in prokaryotic expression system. Methods: Blister fluid and pharyngeal secretions from children with hand-foot-mouth disease in Xi’an area were collected for virus isolation and RT-PCR detection. Enterovirus 71 (EV71) was amplified by reverse transcription-polymerase chain reaction (RT-PCR) Coat protein VP1 gene. The recombinant expression plasmid PQE30 / VP1 was constructed and transformed into E. coli BL21. The VP1 gene was genetically analyzed. Results: The VP1 gene of Xi’an local strain of enterovirus 71 (EV71) was sequenced and compared with that of Fuyang strain (EU913471), indicating that the EV71 isolate in Xi’an of China had a large difference with Fuyang strain. Nucleosides The difference in acid is about 4%. Conclusion: The nucleotide difference of VP1 gene in Xian local strain is more obvious than that in Fuyang strain. This will lay a good foundation for the molecular epidemiology of EV71 in Xi’an and the prevention and control of EV71-induced diseases.