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目的克隆和分析细粒棘球绦虫原头蚴发育调控基因Eg Ral基因。方法根据多房棘球绦虫Em Ral基因序列设计引物,用RT-PCR方法从细粒棘球绦虫原头蚴中克隆Eg Ral基因并将其克隆至pMD19-T载体,经测序后,与线虫、多房棘球绦虫、果蝇和人类等物种Ral基因进行比对分析。结果从细粒棘球绦虫原头蚴cDNA中克隆获得Eg Ral基因,长度为603 bp,编码200个氨基酸,等电点为8.85。经比对,Eg Ral与Em Ral同源性为99%,与线虫、果蝇和人等其他种类Ral基因同源性在47.89%~50.72%之间。进化树分析表明Eg Ral与Em Ral相聚集,与其他物种同源性较低。结论成功从细粒棘球绦虫原头蚴中克隆出Eg Ral基因,其序列具有较高的保守性。
Objective To clone and analyze Eg Ral gene of the developmental regulation gene of Echinococcus granulosus. Methods The primers were designed according to the sequence of Em Ral gene of Echinococcus multilocularis. The Eg Ral gene was cloned from Echinococcus granulosus by RT-PCR and cloned into pMD19-T vector. After sequencing, Echinococcus multilocularis, Drosophila and human Ral genes were analyzed. Results The Eg Ral gene was cloned from cDNA of Echinococcus granulosus, which was 603 bp in length and encoded 200 amino acids with an isoelectric point of 8.85. By comparison, the homology of Eg Ral and Em Ral was 99%, and the homology of other Ral genes with nematodes, flies and humans ranged from 47.89% to 50.72%. Phylogenetic tree analysis showed that Eg Ral and Em Ral clustered with lower homology with other species. Conclusion The Eg Ral gene was successfully cloned from the protoscoleces of Echinococcus granulosus and its sequence was highly conserved.