硕大利什曼原虫无鞭毛体蛋白的基因克隆与序列分析  

CLONING AND SEQUENCE ANALYSIS OF A GENE ENCODING AMASTIN FROM LEISHMANIA MAJOR

作　　者：成军[1] 斯崇文[2] 王勤环[2] 刘妍[1] 钟彦伟[1] 杨继珍[1] 洪卫国[1] 

机构地区：[1]解放军302医院传染病研究所基因治疗研究中心,北京100039 [2]北京医科大学第一医院感染疾病科,北京100034

出　　处：《中国寄生虫学与寄生虫病杂志》2000年第1期30-32,共3页Chinese Journal of Parasitology and Parasitic Diseases

摘　　要：[目的 ]克隆硕大利什曼原虫无鞭毛体蛋白 (amastin)的编码基因序列。 [方法 ]应用核苷酸序列数据库 (GenBank)和表达序列末端片段数据库 (dbEST)的计算机检索与DNA文库的杂交筛选方法。 [结果 ]从dbEST数据库中获得一段 30 9nt的来源于硕大利什曼原虫的基因片段 ,据此设计探针 ,筛选硕大利什曼原虫的DNA文库 ,获得硕大利什曼原虫无鞭毛体蛋白的编码基因。其开放读码框架由 5 5 2个核苷酸组成 ,编码产物由183个氨基酸残基组成。序列分析表明 ,硕大利什曼原虫与锥虫无鞭毛体蛋白一级结构的同源性为 2 3 5 %。 [结论 ]克隆的基因系硕大利什曼原虫表面蛋白编码基因 ,即无鞭毛体蛋白的编码基因。Objective]To clone a gene encoding surface protein from Leishmania major .[Methods] Using T cruzi amastin DNA sequence as a reference,computer search was done on GenBank and dbEST databases by using BLAST path. A Leishmania major DNA library has been constructed and screened by in situ colony hybridization.[Results] A 309nt DNA fragment from Leishmania major was found in dbEST. Leishmania major DNA library was screened using specific primers synthesized according to 309 nt DNA sequence, and a full length coding sequence for Leishmania major amastin was cloned. The coding sequence consisted of 552 nt, and translated into 183 amino acid residues. The homology is 23 5% at amino acid sequence level between Leishmania major and T cruzi amastins. [Conclusion] A full length amastin coding gene for Leishmania major has been cloned.

关 键 词：利什曼原虫 无鞭毛体 基因文库 基因克隆 

分 类 号：R382.22[医药卫生—医学寄生虫学]