细粒棘球绦虫TSP33基因的克隆、生物信息学及表达分析

Cloning,Bioinformatics and Expression Analysis of TSP33 Gene of Echinococcus granulosus

旨在分析细粒棘球绦虫TSP33基因的结构及其在细粒棘球绦虫不同发育阶段的表达情况,预测其作为细粒棘球绦虫诊断抗原的潜在价值。从GenBank中获得Eg-TSP33的cDNA序列并设计特异性引物,提取原头蚴总RNA,反转录cDNA,以cDNA为模板进行PCR扩增,将PCR产物克隆到大肠埃希菌中,测序并进行生物信息学分析。TSP33 cDNA全长864 bp,编码277个氨基酸,Eg-TSP33的预测分子质量为32.11 ku,预测等电点为8.62。RT-PCR结果显示,Eg-TSP33在细粒棘球绦虫成虫、包囊壁及原头蚴中均有表达,且在包囊壁中表达量较多。结果说明,Eg-TSP33可能在细粒棘球绦虫发育过程中可能具有重要作用,为Eg-TSP33分子的诊断价值研究奠定了基础。

The purpose of this study was to analyze the structure of TSP33 gene of Echinococcus granulosus and its expressions in different developmental stages of E.granulosus,and to predict its potential value as a diagnostic antigen of E.granulosus.The cDNA sequence of Eg-TSP33 was obtained from GenBank and the specific primers were designed.The total RNA of the protoscolex was extracted,the cDNA was reversely transcribed,the PCR was carried out using cDNA as a template,and the PCR product was cloned into E.coli and sequenced for biological information analysis.The TSP33 cDNA is 864 bp in length and encodes 277 amino acids.The predicted molecular weight of Eg-TSP33 is 32.11 ku,and the predicted isoelectric point is 8.62.The results of RT-PCR showed that Eg-TSP33 was expressed in the adult,cystic wall and protoscolex of E.granulosus,and it was expressed more in the wall of the cyst.The results indicated that Eg-TSP33 may play an important role in the development of E.granulosus,which laid a foundation for the diagnostic value of Eg-TSP33 molecules.