恶性疟原虫裂殖子表面蛋白1羧基端编码基因真核表达重组克隆的构建

目的构建以恶性疟原虫红内期重要的疫苗候选抗原——裂殖子表面蛋白１（ＭＳＰ１）羧基端编码分子量４２０００蛋白的基因片段为外源基因的可用作候选核酸疫苗的真核表达载体。方法目前对疟疾核酸疫苗的研究仅见于鼠疟，将恶性疟原虫ＦＵＰ株裂殖子表面蛋白１羧基端编码４２０００蛋白的基因片段用常规分子克隆方法，分别克隆入非分泌性真核表达载体ＶＲ１０１２和改建后的分泌型载体ＶＲ１０１２／ＴＰＡ中，通过ＰＣＲ和酶切鉴定出重组克隆。结果成功地构建了真核表达载体ＶＲ１０１２／ＭＳＰ１－４２和ＶＲ１０１２／ＴＰＡ／ＭＳＰ１－４２。结论目前对疟疾核酸疫苗的研究仅见于鼠疟红外期，该研究对研制有效的恶性疟原虫红内期核酸疫苗是一个有益的尝试。其免疫保护作用待进一步研究。

Construction of a couple of eukaryotic expression recombinants containing the gene fragment of 42kD C terminal region of Plasmodium falciparum merozoite surface protein 1

Objectives Nucleic acid vaccine has several major advantages over other types of available vaccines,such as in situ eukaryotic expression and generation of extensively cellular response.To construct a couple of eukaryotic expression recombinants containing the gene fragment of C terminal region (coding for 42kD protein) of Plasmodium falciparum(P.f) merozoite surface protein 1(MSP1),which could be a candidate nucleic acid vaccine. Methods The gene fragment of C terminal region (coding for 42kD protein) of MSP1 from P.f FUP strain was cloned into eukaryotic expression plasmid VR1012 for intracellular expression and VR1012/TPA for secretion by commonly used molecular clone methods.The eukaryotic expression recombinants were identified by PCR and restriction enzyme.Results The recombinants VR1012/MSP1 42 and VR1012/TPA/MSP1 42 were successfully constructed.Conclusions Up to now only nucleic acid vaccine of rodent malaria was reported.This study provides a good basis for us to find out an effective P.f erythrocytic stage nucleic acid vaccine .The protective immune response to these vaccines will be studied in the future.