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目的从新疆株细粒棘球绦虫原头蚴中克隆PDZ结构域蛋白(EgPDZ),进行序列测定和生物信息学分析。方法设计EgPDZ基因特异性引物,用RT-PCR方法从细粒棘球绦虫原头蚴RNA中克隆EgPDZ基因并将其克隆至pMD18-T载体,测序确定序列并进行生物信息学分析。结果 RT-PCR扩增出一长度约600 bp的基因,测序显示其长度为627 bp,编码208个氨基酸,等电点为4.76,为一新基因,命名为EgPDZ。SMART功能分析预测EgPDZ蛋白16～88氨基酸为PDZ结构域。同源性比较表明,EgPDZ与多房棘球绦虫EmPDZ同源性为97.60%,与血吸虫和人等其他种类PDZ基因同源性在18.16%～26.42%之间,而PDZ结构域的序列相似度为42.47%～98.63%。进化树分析表明EgPDZ与EmPDZ相聚集,与其他物种同源性较低。结论成功克隆出细粒棘球绦虫EgPDZ新基因,为进一步研究该基因在细粒棘球绦虫与宿主相互作用中的功能奠定基础。
Objective To clone the PDZ domain protein (EgPDZ) from Echinococcus granulosus of Xinjiang strain for sequence analysis and bioinformatics analysis. Methods The EgPDZ gene-specific primers were designed. The EgPDZ gene was cloned from the sporozoite RNA of Echinococcus granulosus by RT-PCR and cloned into pMD18-T vector. The sequence was sequenced and analyzed by bioinformatics analysis. Results A 600 bp DNA fragment was amplified by RT-PCR. The length of the gene was 627 bp and encoded 208 amino acids. The pI was 4.76. The gene was named as EgPDZ. The SMART functional analysis predicts that the amino acids 16 to 88 of the EgPDZ protein are PDZ domains. Homology comparison showed that the homology between EgPDZ and EmPDZ of Echinococcus multilocularis was 97.60% and the homology of other PDZ genes with Schistosoma japonicum and human was between 18.16% and 26.42%, while the sequence similarity of PDZ domain For 42.47% ~ 98.63%. Phylogenetic analysis showed that EgPDZ and EmPDZ clustered with lower homology with other species. Conclusion The new gene of EgPDZ of Echinococcus granulosus was cloned successfully, which laid the foundation for further study on the function of this gene in the interaction between Echinococcus granulosus and host.