阴道毛滴虫铁氧还蛋白基因的克隆和序列分析  

Cloning and Sequence Analysis of Trichomonas vaginalis Ferredoxin Gene

作　　者：谢辉[1] 王雅静[1] 帖超男[1] 毕世樑[2] 刘佩娜[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]四川大学华西第二医院,成都610041

出　　处：《中国寄生虫学与寄生虫病杂志》2005年第1期24-26,共3页Chinese Journal of Parasitology and Parasitic Diseases

摘　　要：目的　构建阴道毛滴虫铁氧还蛋白 (ferredoxin ,Fd)基因重组质粒 ,并进行克隆及序列分析。　方法　根据Fd基因已知序列 ,设计合成 1对引物 ,应用螯合树脂 (Chelex 10 0 )提取基因组DNA ,用聚合酶链反应 (PCR)扩增出Fd基因 ,克隆入pMD 18T载体 ,转化大肠埃希菌JM 10 9感受态细胞 ,经PCR及酶切鉴定、测序。　结果　Fd基因体外扩增产物长度为 3 0 6bp ,重组质粒经PCR及酶切鉴定结果表明获得正确重组子 ,测序结果表明其片段与已知序列吻合。　结论　克隆出阴道毛滴虫Fd基因。Objective To construct a recombinant plasmid containing ferredoxin gene of Trichomonas vaginalis. Methods Total DNA was extracted from Trichomonas vaginslis with Chelex-100 method and used as templates for PCR. Primers were designed based on the published sequence of the ferredoxin gene and used to amplify the Trichomonas vaginalis gene using PCR method. The ferredoxin gene obtained by PCR technique was directionally cloned into plasmid pMD-18T simple vector. The constructed recombinant plasmid was transferred into E.coli JM109. The transformants were screened and identified by PCR and restriction analysis. The DNA sequence of the gene was determined by Sanger’s method. Results The size of amplified ferredoxin gene was 306bp. The correct recombinant plasmid was isolated and confirmed by PCR and restriction analysis. The DNA sequence of cloned gene was the same as the published sequence. Conclusion The ferredoxin gene was successfully amplified and cloned into plasmid pMD-18T simple vector. The cloned ferredoxin gene could be used to produce recombinant protein and for study of its function.

关 键 词：阴道毛滴虫 蛋白基因 克隆 吻合 PCR 序列分析 测序 酶切 增产 引物 

分 类 号：R711.31[医药卫生—妇产科学] R382[医药卫生—临床医学]