杜氏利什曼原虫LACK抗原基因的克隆和表达  

Cloning and expression of LACK gene of Leishmania donovani

作　　者：卜玲毅[1] 胡孝素[1] 敬保迁[2] 王雅静[1] 马莹[1] 

机构地区：[1]四川大学华西医学中心寄生虫学研究室,成都610044 [2]川北医学院微生物学教研室

出　　处：《中国人兽共患病杂志》2002年第5期42-44,共3页Chinese Journal of Zoonoses

基　　金：国家自然科学基金资助 (No 3 9970 667)

摘　　要：目的　在大肠杆菌中高效表达纯化杜氏利什曼原虫的LACK抗原。方法　以本实验室的重组质粒T -LACK为模板 ,PCR扩增得到LACK抗原基因 ,与表达载体pQE31定向重组 ,转化到宿主菌M 15中进行表达和纯化 ,并以SDS -PAGE和Western -blotting进行鉴定。结果　 1 SDS -PAGE电泳检测 ,重组质粒 pQE31-LACK的全菌蛋白没有出现明显的特异表达带 ,而纯化蛋白和包涵体溶解物在 36kDa处均出现了特异的表达带。 2 36kDa的纯化蛋白被抗利什曼原虫鼠血清清晰地识别。结论　得到了高效表达的LACK纯化蛋白 。Aims To clone the LACK gene of Leishmania donovani and express it in Escherichia coli(E coli) Methods A DNA fragment encoding the LACK gene of Leishmania donovani was obtained from the recombinant plasmid T LACK by PCR The gene was cloned into expressed plasmid pQE31,and then was transformed into E coli M15 for expression and purification The expressed protein was analyzed by SDS-PAGE followed by Western blotting and immuno staninig Results 1 We obtained the protein by three methods from E coli M15 transformed with the recombinant plasmid PQE31 LACK The specific protein band with a molecular weight of 36kDa was detected on SDS PAGE both in the purified protein and solution of inclusion body except the whole protein of E coli 2 The purified protein was identified by immuno staining Conclusion Efficient expression of LACK purified protein has been achieved in E coli and the protein has immunocompetence

关 键 词：利什曼原虫 LACK 聚合酶链反应 克隆 表达 免疫印迹 

分 类 号：R382.22[医药卫生—医学寄生虫学]