Echinococcus granulosus钙结合蛋白基因的克隆和序列分析

Cloning and Sequence Analysis of the CABPs Gene from Echinococcus granulosus

目的 获得细粒棘球蚴Calcium bindingprotein(CaBPs)基因序列资料 ,进行序列分析 ,为包虫病免疫预防研究奠定基础。方法 从包囊内获取细粒棘球蚴原头蚴 (protoscolices) ,提取RNA ,逆转录成cDNA ,用特异引物扩增CaBPs基因 ;并将其克隆到T载体后进行序列测定和分析。然后登录Gene/Bank数据库。结果 以cDNA为模板获得 3个核苷酸长度不同的基因 ,分别是CaBP1为 914bp ,CaBP2为 10 95bp ,CaBP3为 10 39bp。同源性比较结果表明 :来自新疆羊源CaBPs基因序列和巴西 (Brazil)羊源基因CaBPs同源性为 90 %以上 ;从cDNA获得的 3个CaBPs基因序列的比较分析说明钙结合蛋白可能存在一个家族 ,有不同的成员组成。结论 CaBPs在E .granulosus虫体发育过程中具有十分重要的功能 ,到目前为止其机理尚不完全明了 ,CaBPs基因的获得为进一步研究其结构和功能 ,以及对包虫病的防治具有重要意义。

Objective To obtain Calcium binding protein(CaBPs)gene of Echinococcus granulosus and compare their homology with those reported nucleotide sequence of CaBPs of E.granulosus.Method CaBPs genes of E.granulosus from the cDNA templates were amplified by using specific primers and were cloned into T vector for sequencing and analyzing.Results Three sequences, 914bp,1095bp and 1039bp, were obtained. The result of homology comparison indicated that the homology is 90% between CaBPs gene sequence from XinJiang and the gene sequence from Brazil and belong to the same gene family. Conclusion Three CaBPs genes have been isolated from E.granulosus protoscoleces. The CaBPs cloned in this study belong to one gene family which can been used as candidate antigen gene to developed vaccine and their biological functions studying.