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
出处
《中国人兽共患病杂志》
CAS
CSCD
北大核心
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)
