关键词: 约氏疟原虫, 裂殖子表面蛋白, 基因重组

Abstract: Objective To construct recombinant plasmid pPGEX-2T-PyMSP1V30 bearing the gene coding for Merozoite surface protein 1 of Plasmodium yoelii and expressed as a soluble protein in Escherichia coli and purified according to good manufacturing practices.The aim is to explore the possiblity of immunogenicity detecting and early specific diagnosis of malaria,and provide the research base in studying on higher efficiency polyvalent gene vaccine anti-erythrocytic stage parasites further.Methods Using polymerase chain reaction(PCR) technique,the Merozoite surface protein 1 was amplified from genomic DNA of Plasmodium yoelii amplifying to a 416 bp of 4855-5271 fragment,then cloned into the plasmid pPGEX-2T,and pyMSP1-17 was constructed.pyMSP1-17 expressed the fusion protein in E.coli DH-5α.Results The recombinant PGEX-2T-PyMSP1-17 contained the MSP1-17 from P.yoelii.The fusion 41 kilodalton protein of GST-MSP1 was collected and purified with the total concentration of 46 mg/L.These recombinants proteins were used to immunize C3H/He mice.The specificity and subtyping of the antibodies from the immunized C3H/He mice was carried out.Furthermore,showed reactivity with P.vivax,P.cynomolgi,P.knowlesi and P.coatneyi schizonts by immunofluorescence assays(IFA) with the titer 1∶400-1∶1000.Conclusion It could highly express fusion protein in E.coli that recombinant Merozoite surface protein 1 of P.yoelii and the fusion protein possessing the immunogenicity and antigenicity.It is a scientific base for malaria early diagnosis study.With in higher production of fusion protein and better immunogenicity it maybe used as an efficient candidate genetic vaccine against the asexual blood stages of plasmodium.