恶性疟原虫FCC1/NH株裂殖子表面蛋白MSP1第2～5区基因的PCR扩增及克隆

PCR AMPLIFICATION AND CLONING OF THE GENG ENCODING BLOCK 2-5 IN MSP1 FROM THE FCC1/HN ISOLATE (SOUTHERN CHINA)OF PLASMODIUM FALCIPARUM

构建恶性疟原虫FCC1／HN株MSP1基因2～5编码区真核表达质粒pcDNA3／MSP1，为FCC1／HN株MSP1的体外表达及其基因工程疫苗的研制奠定基础。方法采用PCR技术对恶性疟原虫海南分离株（FCC1／HN）基因组DNAMSP1第2～5区基因片段进行扩增．扩增产物经纯化后用Xhol和Aaal双酶切然后定向克隆入pcDNA3质粒，转化大肠杆菌TG1．再用相同内切酶酶切和PCR扩增对重组子进行鉴定。结果筛选出编码FCC1／HN株MSP1第2～5区基因片段的真核表达质粒pcDNA3／MSP1。结论编码FCC1／HN株MSPI第2～5区基因片段的真核表达质粒pcDNA3／MSP1的构建，为恶性疟原虫FCC1／HN株MSP1的体外表达及其基因工程疫苗的研制奠定基础。

Aim To express MSP1 protein and develop protein vaccine of malaria,Constructing of the eukaryotic expression plasmid pcDNA3/MSP1. Methods Using polymerase chain reaction(PCR) technique,the gene fragment coding for block 2 -- 5 in MSP1 was amplified from genomic DNA of southern China isolate (FCC1/HN) of P. falciparum. The PCR product was purified and digested with Xhol and Apal,and the generated DNA fragment was cloned into plasrnid pcDNA3,and transformed into Escherichia coli(E. coli) strain TG1.The recombinant plasmids were screened and identified by Xhol and Apal digestion and PCR amplification. Result The result demonstrated that the recombinant plasmid pcDNA3/MSP1 contains the exogenous gene encoding the block 2--5 in MSP1 from FCC1/HN isolate of Plsmodium falciparum. Conclusion These findings are very important for the expression of MSP1 in vitro and the development of malaria vaccine.