以恶性疟原虫裂殖子表面蛋白1的17区片段基因为基础构建的复合核酸疫苗候选质粒

目的 :构建以恶性疟原虫裂殖子表面蛋白 1的 17区基因为基础的复合核酸疫苗。方法 :将恶性疟原虫 FUP株 MSP1的 17区基因片段与包含若干个内源性和外源性 T细胞位点相应的基因片段相连 ,构成复合基因 ( hybrid gene,HG) ,将其分别克隆入非分泌性真核表达载体 [VR10 12和 p CDNA3.1( - ) ]和经改建后的分泌型载体 VR10 12 /TPA中 ,通过 PCR和酶切鉴定重组克隆。结果和结论 :经鉴定复合基因正确地克隆入真核表达载体 ,成功地构建了 VR10 12 /HG,p CD-NA3.1/HG和 VR10 12 /TPA/HG。结论 :为探讨疟疾核酸疫苗的效果及 T细胞位点的作用打下了基础。

CONSTRUCTION OF HYBRID NUCLEIC ACID VACCINES BASED ON PLASMODIUM FALCIPARUM MEROZOITE SURFACE PROTEIN1BLOCK17REGION

AIM:To construct hybrid nucleic acid vaccines which contain Plasmodium falciparum merozoite surface protein 1 (MSP1 ) block 17 gene and gene fragment coding for several T cell epitopes. METHODS:MSP1 block 17 gene of FUP strain was connected with a polyvalent gene fragment which contains several T cell epitopes from MSA1 ,MSA2 ,RESA,IL- 1 and Tetanus toxin, the connected gene fragment (hybrid gene,HG) was cloned into eukaryotic expression plasmid pCDNA3.1 (- ) ,VR1012 for intracellular expression,VR1012/TPA for secretion, the recombinants were identified by PCR and restriction enzyme digestion. RESULTS& CONCLUSION: The hybrid nucieic acid vaccines: VR 1012/HG, VR 1012/TPA/HG and pCDNA 3.1/HG were successfully constructed.