阴道毛滴虫Rac1蛋白的cDNA克隆和序列分析(英文)  

Molecular Cloning and Characterization of a Rac1 Homologue cDNA from Trichomonas vaginalis

作　　者：傅玉才[1] 章家新[1] 郑晓虹[1] 刘红[1] 

机构地区：[1]汕头大学医学院寄生虫学教研室,汕头515031

出　　处：《中国寄生虫学与寄生虫病杂志》2004年第5期290-293,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：汕头大学研究与发展基金项目 (No.L0 0 0 1 3)~~

摘　　要：目的　获得阴道毛滴虫Rac1蛋白的cDNA克隆 ,研究其在细胞周期中的调解作用。　方法　提取阴道毛滴虫总RNA ,构建cDNA表达文库 ,随机分离cDNA克隆并测序。用在线生物分析软件NCBIBLAST、ClustalW以及Treeview等程序进行序列分析。　结果　获得一株有 714bp的cDNA克隆。序列分析表明 ,该克隆开放阅读框具 60 0bp ,推测肽链具 2 0 0个氨基酸。该肽链与Rho家族中Rac1鸟苷三磷酸 (GTP)酶同源性最高 (>60 % ) ,并具多种RhoGTP酶的保守基序 ,如GTP结合部位、GTP酶激活蛋白作用基序、GTP分离抑制因子作用基序、鸟嘌呤核苷酸交换因子作用基序等。进化树分析显示该克隆属于Rac亚家族GTP酶 ,与原虫Rac1蛋白最接近。　结论　该克隆属RhoGTP酶的Rac亚家族 ,很可能是阴道毛滴虫的Rac1蛋白。Objective To clone and characterize a Rac1 homologue from Trichomonas vaginalis for studying cell cycle of the organism. Methods A cDNA library derived from T. vaginalis mRNA was constructed into λ TriplEx2 phage vector. An expression sequence tag program was launched. Sequences of cDNA clones were analyzed using NCBI BLAST algorithms, and ClustalW and Treeview programs. Results A cDNA clone with a length of 714 base pairs was isolated. The sequence analysis showed that the cDNA clone has an open reading frame with 600 bp. The deduced amino acid sequence from the open reading frame contains 200 residuals and is most homologous to Rac1 subfamily of Rho GTPases with >60% identity. The conserved sequence elements of Rho GTPases, such as GTP binding sites, GTPase activating protein (GAP) interaction motifs, GTP dissociation inhibitors (GDI) interaction motifs, guanine nucleotide exchange factor (GEF) interaction elements, etc, were detected in the amino acid sequence. The phylogenetic analysis showed that the cDNA clone is grouped in the Rac subfamily and is more closely related to Rac1 proteins of protozoa. Conclusion The cDNA clone isolated belongs to Rac subfamily of Rho GTPases and is probably a Rac1 protein of T. vaginalis.

关 键 词：阴道毛滴虫 L蛋白 CDNA克隆 序列分析 GTP酶 作用 酶激活 肽链 NCBI 生物分析 

分 类 号：R373.21[医药卫生—病原生物学] R135.12[医药卫生—基础医学]