日本血吸虫微管蛋白基因的分离和cDNA序列分析  

An Isolation and Sequence Analysis of the cDNA Encoding Schistosoma japonicum Beta-tubulin

作　　者：刘传爱[1] 肖建华[1] 梁瑜[1] 廖力[1] 张愉快[1] 曾桥[1] 

机构地区：[1]南华大学病原生物学实验中心,湖南衡阳421001

出　　处：《实用预防医学》2005年第1期20-22,共3页Practical Preventive Medicine

基　　金：湖南省教育厅重点项目 (No.0 3A0 4 1 )

摘　　要：目的　分离和鉴定日本血吸虫 (Schistosomajaponicum ,Sj)新基因 ,为防治日本血吸虫病提供药物靶标或候选疫苗。　方法　构建日本血吸虫成虫cDNA文库 ,随机挑取重组阳性克隆进行测序 ,对部分序列进行引物步移法测序 ,获取其全长cDNA序列 ;采用生物信息学等技术对该cDNA序列进行开放阅读框 (ORF)的寻找、编码氨基酸的推导、核苷酸和氨基酸同源性比较以及蛋白质二级结构的初步预测。　结果　获得了 1个日本血吸虫新基因—微管蛋白基因 ,全长14 3 9bp ,编码 44 3个氨基酸 ,与肝片形吸虫微管蛋白基因具有 79%的同源性。编码蛋白的理论分子量为 7.2 198KDa ,等电点为 9.12 ;抗原表位可能位于cDNA序列 3 73～ 3 96处。　结论　获得了日本血吸虫微管蛋白基因的全长cDNA序列 ,为该基因功能的实验性鉴定工作奠定基础。Objective To isolate and identify novel genes from an adult Schistosoma japonicum (Sj) cDNA library, and find new drug targets or vaccine candidates for preventing, controlling and treating schistosomasis. Methods A cDNA library was constructed from Sj (Chinese strain) adult stage. Clones were selected randomly from the cDNA library and were sequenced by using the method of expression sequence Tags (ESTs). Novel genes were acquired by primer-walking. By using bioinformatics,the cDNA sequence encoding Sj beta-tubulin was analyzed, including searching the ORF ,translating the nucleotide to protein sequence ,similarity searches and secondary structure predication. Results Beta-tubulin genes, 1 439 bp and coding for 443 amino acids, was acquired and was submitted to GenBank and to get an accession number that was AY220457. The beta-tubulin gene of S. japonicum and Fasciola hepatica were homologous with an identity of 79%. The academic pI is 9.12, and the antigen determinant is probably from 373 to 396 on the cDNA sequence. Conclusion The full-length cDNA sequence encoding Sj beta-tubulin was obtained, which gave a basis for further functional study of this gene.

关 键 词：日本血吸虫 微管蛋白 序列分析 表达序列标签 引物步移法 

分 类 号：R383.24[医药卫生—医学寄生虫学]