【摘要】 【目的】建立日本血吸虫 (Schistosomajaponicum ,Sj)表达基因的大规模随机测序体系。【方法】从SjcDNA文库中随机挑选出单个重组克隆 ,扩增出cDNA插入片段 ,采用PCR直接序列测定方法对插入片段 5′端进行部分序列测定。通过互联网将获得的表达序列标签 (expressedsequencetags ,EST)送入WHO血吸虫基因库进行同源性检索。【结果】完成了 16 0个Sj表达基因EST序列测定 ,获得了 12 6个可用于分析的EST ,并在GenBank登录。 10个为Sj已知编码基因 ,其中 3个为Sj抗原基因全长cDNA。【结论】建立了Sj表达基因EST测序及同源分析体系 ,为进一步开展Sj表达基因的研究奠定了基础。更多还原

【Abstract】 Objective To establish large-scale sequencing system for expressed genes of S.japonicum(S j). Methods The cDNA clones were selected randomly from S j cDNA library, and the insert fragments were amplified by PCR. The expressed sequence tags (ESTs) were obtained by PCR directed sequencing from 5′ end of the partial sequence, then the ESTs were compared with all sequences in WHO Schistosoma GenBank data bases. The ESTs were submitted to NCBI dbEST for obtaining accession numbers. Results 160 recombinant clones were picked from the cDNA, and the inserts of cDNA were identified by PCR. 126 ESTs were generated by PCR directed sequencing. 10 ESTs belonged to “identified genes of Schistosoma” for they matched with Schistosome sequences in the databases, among them, 3 ESTS contain full cDNA sequences with ORF. Conclusion The system for large scale sequencing of Chinese strain Schistosoma japonicum is successfully established and provides the basis for further study related to S j expressed genes.更多还原