摘要
目的 运用表达序列标签 (EST)技术 ,从日本血吸虫 (Schistosomajaponicum ,Sj)cDNA文库中分离、鉴定出有潜在应用价值的血吸虫基因序列。方法 应用EST方法 ,从日本血吸虫cDNA文库中随机挑选出单个重组克隆 ,PCR扩增出插入片段 ,并进行PCR产物直接测序 ;将获得的序列资料与EBI和GenBank进行BLASTn和BLASTx同源性检索 ,对可能的酶类基因序列进行分析。结论 采用EST策略 ,可获得日本血吸虫 (中国大陆株 )若干酶类基因序列EST ,为进一步的cDNA全长扩增和克隆奠定基础。
Aim To isolate and identify Schistosoma japonicum(Chinese strain)valuable genes from a directional cDNA library by EST strategy.Methods The cDNA clones were selected randomly,and the ESTs were obtained by PCR directed sequencing,then the novel ESTs were submitted to NCBI dbEST and GenBank for obtaining accession numbers.Results 362 recombinant clones picked out of the cDNA library,and the inserts of cDNA clones were identified by PCR.90 ESTs were generated by PCR directed sequencing.Among them,7 genes associated with enzymes were studied further.Conclusion This study shows that EST strategy can be used to discover novel valuable genes of Schistosoma japonicum.
出处
《中国人兽共患病杂志》
CSCD
北大核心
2001年第4期46-48,共3页
Chinese Journal of Zoonoses
基金
中山医科大学"2 11"工程重点学科资金 (No.9816 9)
国家自然科学基金资助 (NO .30 0 70 6 83)
关键词
日本血吸虫
中国大陆株
CDNA文库
表达序列标签
酶
糖酵解酶
Schistosoma japonicum(chinese strain)
cDNA library
Expressed Sequence Tags(EST)
Enzyme
Glycolytic enzyme
