摘要
目的对细粒棘球蚴(E.granulosus)翻译延伸因子(translationelongationfactor)EF-1基因片段进行分子克隆及序列分析。方法从细粒棘球蚴原头蚴中提取RNA,采用RT-PCR技术扩增出细粒棘球蚴EF-1基因片段,与pGEM-T载体重组并转化E.ColiJM109,经筛选扩增获得重组基因克隆,再进行序列测定和分析。结果成功构建了EF-1/pGEM-T/JM109克隆体系,测序表明该基因开放阅读框为693bp,与已发表基因核苷酸序列相比,同源性为99%,推导编码氨基酸序列同源性为99%。结论扩增获得的基因片段可能为细粒棘球蚴(E.granu-losus)翻译延伸因子EF-1基因片段。
Objective To obtain and analyze sequence EF- 1 gene, and lay bases for screening candidate antigen of echinococcus granulosus. Methods Total RNA was extracted from protoseoles of cysts from humam. The specific primers were designed according to published nucletid sequence in the genbank database. The EF - 1 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 693bp has been amplified successfully by RT-PCR. Comparision of the DNA and amino acid sequence deduced from cDNA with the published EF- 1 gene sequence of echinococcus granulosus in the genbank revealed 99 % identity samely. Conclusion Because the EF - 1 play an importante role in energy metabolism of E. granulosus, EF- 1 gene gained from protoscoles can be used as candidate antigene gene to develope vaccine and its biological functions studying.
出处
《宁夏医学杂志》
CAS
2005年第8期510-513,共4页
Ningxia Medical Journal
基金
国家自然科学基金项目(项目编号:30260105)
关键词
棘球蚴病
基因
结构
克隆
分子
序列分析
Echinococcosis
Eenes structural
Clonging molecular
Sequencing
