阴道毛滴虫黏附蛋白65-3基因的cDNA克隆与序列分析  

Cloning and Sequence Analysis of Trichomonas vaginalis Adhension Protein 65-3

作　　者：朱晓燕[1] 王雅静[1] 杨树国[2] 毕世樑[3] 廖琳[1] 

机构地区：[1]四川大学华西基础医学与法医学院寄生虫学教研室,成都610044 [2]郧阳医学院寄生虫学教研室,十堰442000 [3]四川大学华西第二医院妇产科,成都610041

出　　处：《热带医学杂志》2007年第8期715-717,725,共4页Journal of Tropical Medicine

基　　金：国家自然科学基金(No.39970667)

摘　　要：目的对阴道毛滴虫黏附蛋白65-3基因进行cDNA克隆与序列分析。方法Trizol试剂提取阴道毛滴虫基因组RNA。以总RNA为模板,RT-PCR扩增黏附蛋白65-3基因,定向克隆入pMD-18T线形载体,构建pMD-18T-ap65-3重组质粒,并进双行酶切﹑PCR鉴定及测序分析。结果RT-PCR扩增出阴道毛滴虫黏附蛋白65-3基因。经凝胶电泳﹑PCR鉴定和限制酶切鉴定,成功的构建出pMD-18T-ap65-3重组质粒。序列分析表明,黏附蛋白65-3基因开放阅读框长为1 704 bp,与GenBank上公布的黏附蛋白65-3基因序列比较,其同源性高达99.6%。结论阴道毛滴虫黏附蛋白65-3基因体外扩增及测序的成功,为进一步研究阴道毛滴虫的致病机理及滴虫病防治方法奠定了基础。Objective To clone and sequence the cDNA coding for the adhensin protein 65-3 of Trichomonas vaginalis （ap65-3 gene）. Method Total RNA was extacted from Trichomonas vaginalis using Trizol reagent. The ap65-3 gene amplified by RT-PCR was cloned into pMD-18T vector. The recombinant plasmid pMD-18T-ap65-3 was identified by PCR and restriction analysis, and the inserted fragment was sequenced. Result The ap65-3 gene was amplified from total RNA by RT-PCR. The recombinat plasmid pMD-18T-ap65-3 was constructed and comfirmed by PCR and restriction analysis. The sequence analysis revealed that ap65-3 gene contains an open reading frame with 1 740 bp and has 99.6% homology with the sequence of ap65-3 gene published on GenBank. Conclusion The ap65-3 gene was cloned and sequcenced succesfullly. The gene can be used in the future analysis of the pathogenic mechanisms of Trichornonas vaginalis and for the development of treatment for trichomoniasis.

关 键 词：阴道毛滴虫 黏附蛋白65-3 基因克隆 序列分析 

分 类 号：R382.21[医药卫生—医学寄生虫学]