摘要
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 and characterize EgPDZ,which is similar to the PSD-95,Dlg,and ZO-1（PDZ） domains,from Echinococcus granulosus（Eg） protoscolices and analyze EgPDZ using bioinformatics.Methods Specific primers for EgPDZ were designed and the EgPDZ gene was amplified by RT-PCR from Eg protoscolex RNA.Then,EgPDZ gene fragments were cloned into a pMD18-T vector for sequencing and analysis using bioinformatics.Results The PCR products of EgPDZ were about 600 bp and were identified as a new gene by sequence analysis.This gene,designated EgPDZ,had a length of 627 bp and encodes 208 amino acid with a PI of 4.76.SMART analysis predicted that amino acids 16 to 88 of the EgPDZ protein represent a PDZ domain.Bioinformatics indicated that the EgPDZ sequence has 97.60% homology with EmPDZ and 18.16%-26.42% homology with the PDZ genes of other species such as schistosomes and human beings.However,the PDZ domain sequence of EgPDZ gene has a higher homology with that of other species of about 42.47%-98.63%.Phylogenetic analysis showed that EgPDZ was clustered with EmPDZ and had a lower homology with the PDZ genes of other species.Conclusion A new EgPDZ gene was cloned from protoscolices of Eg,providing a foundation for further study of the role of EgPDZ in host-Eg interaction.
出处
《中国病原生物学杂志》
CSCD
2010年第8期590-593,共4页
Journal of Pathogen Biology
基金
国家自然科学基金资助项目(No.30960341
30760228)
新疆维吾尔自治区科技支疆项目(No.200891126)
新疆维吾尔自治区包虫病基础医学重点实验室开放课题项目(No.XJDX0202-2008-03)
