摘要
目的 从日本血吸虫成虫cDNA文库中分离、鉴定和分析全长cDNA序列 ,为日本血吸虫病的防治提供候选疫苗和药物靶点。方法 从构建日本血吸虫成虫cDNA文库中挑取重组阳性克隆进行 5’测序获取表达序列标签 (ExpressedSe quenceTag ,EST) ,在此基础上进行引物延伸法 (primerextension)测序 ,直至获取全长cDNA序列 ,将获取的基因登录Gen Bank ,并进行生物信息学分析。结果 获得 7个日本血吸虫全长cDNA序列 (GenBank登录号分别为AY336 4 93～AY336 4 99) ,其中 6个为日本血吸虫新基因 :类转录因子BTF3(similartotranscriptionfactorBTF3)、延长因子 1-α(elongationfactor 1-alpha ,EF1-α)、富含谷氨酰胺四联重复肽 (smallglutamine-richtetratricopeptide ,TRP)、鸟氨酸氨基转移酶 (ornithineaminotransferase,OA)、内皮差异相关因子 1(endothelialdifferentiation -relatedfactor 1,EDF - 1)和类基因转录增强因子 (similartotestisenhancedgenetranscriptprotein ,TEGT) ;另 1个是精氨酸酶 (arginase)基因 ,比原发现的基因序列长 4 5 6bp。生物信息学分析发现类基因转录增强因子为跨膜蛋白 ,具有 6个跨膜区 ;各基因与人、鼠、蟾和曼氏血吸虫等物种的基因有 37%～ 85 %的同源性 ;
To acquire and analyze full length cDNAs of adult S. japonicum (Chinese strain) from an adult S. japonicum cDNA library and to search new vaccine candidates and drug targets for prevention,controls and treatment of schistosomasis.Positive clones were selected from an adult S. japonicum (Chinese strain) cDNA library and were sequenced by expressed sequence tags (EST) strategy and primer extension method. All genes were analyzed in GenBank databases by BLAST programs and other programs submitted to GenBank. The results showed that 7 full length cDNAs were acquired and submitted to GenBank with accession numbers of AY336493～AY336499. Six of them were new genes of S. japonicum that were similar to transcription factor (BTF3), elongation factor 1 alpha (EF1 α), small glutamine rich tetratricopeptide (TRP), ornithine aminotransferase (OA), endothelial differentiation related factor 1 (EDF 1) and similar to testis enhanced gene transcript protein (TEGT). One of them were an update gene that was arginase. TEGT protein was a transmembrane protein that had 6 transmembrane helixs. All genes were homology to sequences of homo sapiens, Mus musculus, Xenopus laevis and Schistosoma mansoni with 36～85% identity. There were many motifs or domains in the genes that predicted the genes had related functions, such as NAC domain, arginase domain, TEF domain, TRP domain, aminotransferase domain, HTH XRE domain and NAF domain. It is concluded that EST stratagy and primer extension methods are rapid, efficient and economical methods to obtain the full length cDNAs of the adult stage Schistosoma japonicum (Chinese strain).
出处
《中国人兽共患病杂志》
CSCD
北大核心
2004年第12期1058-1061,共4页
Chinese Journal of Zoonoses
基金
湖南省卫生厅重点项目基金的资助 (NO .2 0 0 1-Z0 5 )
关键词
日本血吸虫
CDNA文库
基因
生物信息学
Schistosoma japonicum(Chinese strain)
cDNA library
gene
bioinformatics
