恶性疟原虫裂殖子表面抗原2基因片段的克隆及表达

CLONING AND EXPRESSION OF MEROZOITE SURFACE ANTIGEN 2 FROM PLASMODIUM FALCIPARUM

目的 构建恶性疟原虫海南分离株 (FCC- 1/ HN)裂殖子表面抗原 2 (MSA2 )基因片段的重组真核表达质粒p BK/ MSA2。 方法 采用限制性内切酶法从重组的大肠杆菌 -分枝杆菌穿梭质粒 p BCG/ MSA2中分离出经过测序鉴定的 MSA2基因片段 ,将其亚克隆入真核表达载体 p BK- CMV,构建重组真核表达质粒 p BK/ MSA2。经 IPTG诱导 ,重组质粒在大肠杆菌 DH5 α中进行表达 ,并进行十二烷基磺酸钠 -聚丙烯酰胺凝胶电泳 (SDS- PAGE)及免疫印迹 (Western- blot)分析。 结果 从 p BCG/ MSA2中分离出 MSA2基因片段 ,成功构建 p BK/ MSA2重组质粒 ;SDS- PAGE及 Western- blot分析结果显示 ,特异性蛋白条带的分子质量约为 31ku。 结论 恶性疟原虫

Objective To construct eukaryotic expression recombinant plasmid of merozoite surface antigen 2(MSA2) of FCC 1/HN isolate of Plasmodium falciparum . Methods Using restriction enzymes, the sequenced and identified MSA2 gene fragment was isolated from the recombinant E.coli Mycobacteria shuttle plasmid pBCG/ MSA2, then subcloned into eukaryotic expression vector pBK CMV, and recombinant eukaryotic expression plasmid pBK/MSA2 was constructed. Induced by IPTG, the recombinant plasmid was expressed in E.coli DH5α, and its expressing product was identified by SDS PAGE and Western blot. Results The MSA2 gene fragment was isolated from pBCG/MSA2, and a recombinant plasmid pBK/MSA2 was successfully constructed. The results of SDS PAGE and Western blot revealed that the molecular weight of recombinant protein was approximately 31 ku and can be specially recognized by positive serum of malarial patients from Hainan Province. Conclusion The gene encoding MSA2 from P. falciparum FCC 1/HN strain was successfully expressed in E.coli DH5α.