日本血吸虫睾丸增强转录蛋白样蛋白基因的获得和分析  

Aquiring and Analyzing of Testis-enhanced Transcript Protein-like Protein Gene of Schistosoma Japonicum

作　　者：张愉快[1] 肖建华[1] 刘彦[1] 曾桥[1] 汪志刚[1] 曾谷清[1] 

机构地区：[1]南华大学病原生物学研究所,中国湖南衡阳421001

出　　处：《实用预防医学》2004年第4期661-664,共4页Practical Preventive Medicine

基　　金：湖南省卫生厅重点项目 (No.2 0 0 1 -Z0 5) ;教育厅科研基金资助课题 (4- 0 2 -JY - 0 2 -C388)

摘　　要：目的　从日本血吸虫 (Schistosomajaponicum ,Sj)成虫cDNA文库中获得并分析日本血吸虫新的蛋白基因 ,为日本血吸虫病的防治提供侯选疫苗和药物靶点。　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 ,对部分序列进行步移法测序获取全长cDNA ,登录GenBank ,利用BLAST程序对同源性高的基因进行核苷酸和氨基酸水平的同源性比较 ;并利用 pcgene软件对其蛋白质结构进行初步分析。　 结果　获得了 1个日本血吸虫新基因睾丸增强转录蛋白样蛋白 (testis -enhancedtranscriptprotein -likeprotein ,AY3 3 64 99) ,长 10 5 3bp ,编码 2 19个氨基酸 ,与睾丸增强转录蛋白样蛋白具有 5 5 %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　步移法测序和生物信息学技术相结合有利于发现日本血吸虫新基因。Objective To acquire and analyze adult stage Schistosoma japonicum (Sj) new genes from an adult Schistosoma japonicum cDNA library,and to search new vaccine candidates and drug targets for preventing,controlling and treating schistosomiasis. Methods A cDNA library was constructed from Schistosoma japonicum (Chinese strain) adult stage. Clones were selected randomly from the cDNA library and were sequenced. Novel genes were acquired after analysis in GenBank databases by BLAST programs and other programs,and the analysis of protein was carried out with pcgenge software. Results ubiquitin-conjugating enzyme E gene,1053 base pair and coding for 219 amino acids,was acquired and was submitted to GenBank and got an accession number of AY251608. This gene was homologous with human ubiquitin-conjugating enzyme E gene with identity 55% ;The academic pI is 9.12;and the antigen determinant is probably from 373 to 396 on the cDNA sequence. Conclusion It is a feasible method to discover and acquire novel genes of Schistosoma japonicum by combination of Primer-walking sequencing and Bioinformatics technique.

关 键 词：日本血吸虫 CDNA文库 步移法 生物信息学 睾丸增强转录蛋白样蛋白 基因 

分 类 号：R383.24[医药卫生—医学寄生虫学]