日本血吸虫TEGT基因的cDNA克隆和生物信息学分析

Cloning and Sequence analysis of a cDNA clone isolated from Schistosoma japonicum

目的克隆日本血吸虫新基因TEGT(Sj.TEGT)的全长序列。方法根据编码日本血吸虫大陆株的EST基因片段的序列,设计5’RACE的系列引物,扩增Sj.TEGT的5’端,测序后预测该基因的全长ORF;再以此设计引物,以日本血吸虫中国大陆株成虫mRNA为模板,用RT-PCR方法扩增得到Sj.TEGT的全长cDNA。结果从日本血吸虫中国大陆株成虫mRNA中克隆到1个TEGT相关基因,命名为Sj.TEGT基因。其编码的蛋白的分子质量大约为30ku;对其编码的氨基酸序列的分析显示其编码蛋白有6个跨膜螺旋区。结论Sj.TEGT是一个新基因,有必要对其功能进行深入研究。

Objective To obtain full length the eDNA from Schistosomajoponicum. Methods The total KNA extracted from Schistosoma joponicum was taken as template for reverse transcription. With the method of PACE and RT-PCR. technique, we obtained a clone of the full length eDNA with similar to testis-enhanced gene transcript （TEGT）, so it was named Sj.TEGT of Schistosoma joponicum. Results About 60bp belong 5＇end was successfully cloned ,the new gene represents a TEGT gene with calculated molecular masses of 30ku.The deduced amino acid sequence analyse showed 6 trammembrane structure. Conclusion The novel gene Sj.TEGT needs further study.