细粒棘球蚴中国大陆株zw-5基因的克隆和生物信息学分析

目的获得细粒棘球蚴zw-5基因序列，进行序列分析，为研究包虫病疫苗候选基因作好前期工作。方法从包虫病患者体内获取细粒棘球蚴原头蚴提取总RNA，根据国外细粒棘球蚴Eg-CWGRS-12D11基因的已知序列设计一对引物，采用RT—PCR技术扩增，将PCR产物纯化后，将其克隆到pGEM—T载体后进行序列测定以及生物信息学分析。结果用RT—PCR成功扩增出细粒棘球蚴中国大陆株zw-5基因，测序表明该基因开放阅读框为624bp，与已发表基因核苷酸序列相比，同源性为99％。结论在国内首次获得细粒棘球蚴zw-5基因序列，对进一步研究其结构和功能以及包虫病的防治具有重要意义。

Cloning and Bioinformatics Analyzing of the zw-5 Gene From Echinococcus Granulosus of China

Objective To obtain and analyze sequence zw-5 gene,and lay bases for screening candidate antigen of Echinococcus granulosus. Methods Total RNA was extracted from protoscoles of cysts from humam origin. The specific primers were designed according to published nucleotid sequence in the Genbank database.The zw-5 gene of Echinococcus granulosus was amplified by RT-PCR and cloned into pGEM-T vector for sequencing and analyzing.Results A cDNA sequence with an open reading frame of 624bp has been amplified successfully by RT-PCR .Comparision of the DNA and amino acid sequence deduced from cDNA with the published zw-5 gene sequence of Echinococcus granulosus in the Genbank revealed 99% identity samely.Conclusion Zw-5 gene gained in China from protoscoles can be used as candidate antigen gene to develop vaccine and its biological functions studying.