【摘要】 目的　构建恶性疟原虫海南分离株 ( FCC1/ HN)裂殖子表面蛋白 2 ( MSP2 )融合 HBs Ag基因片断真核表达质粒 p VX-ORF1- Pf MSP2 - HBS及重组真核表达质粒 p VXORF1- Pf MSP2 ,为疟疾核酸疫苗及蛋白疫苗的研制奠定基础。　方法　 ( 1)采用 PCR技术对恶性疟原虫 FCC1/ HN株 MSP2 原核表达质粒 PET2 8α- MSP2 的 MSP2 基因进行扩增 ,扩增产物经纯化后用 Bam H 酶切 ;质粒 p VXORF1- Pv MSP1 .1 9- HBS用相同的酶酶切 ,经纯化后用 T4DNA连接酶将其与酶切后纯化的 MSP2 基因连接 ;( 2 )分别用Bam H +Xho 双酶切 PET2 8α- MSP2 质粒 DNA和 p VXORF1质粒 DNA,纯化后将目的基因片断用 T4DNA连接酶连接 ;将 ( 1)、( 2 )连接产物分别转化大肠杆菌 DH5α。于氨苄青霉素阳性 L B培养平板上筛选阳性克隆 ,酶切电泳鉴定。　结果　筛选出的重组子为编码 FCC1/ HN MSP2 基因片断的重组质粒 p VXORF1- Pf MSP2 - HBS及 p VXORF1- Pf MSP2 。　结论　编码 FCC1/ HN MSP2 基因片断真核表达质粒 p VXORF1- Pf MSP2 - HBS和 p VXORF1- Pf MSP2 的构建为疟疾核酸疫苗及蛋白疫苗的研制奠定了基础。更多还原

【Abstract】 Objective To construct recombinant plasmid of the southern China FCC1/HN isolate of Plasmodium falciparum bearing the gene en-coding for hybid merozoite surface protein 2-Hepatitis B virus surface protein fusion pVXORF1-pfMSP 2-HBS and eukaryotic expression recombinant plasmid pVXORF1-pfMSP 2. Methods (1) Using polymerase chain reaction (PCR) technique, the MSP 2 gene was amplified from genomic DNA of prokaryotic expression recombinant plasmid PET28α-MSP 2 of the southern China isolate (FCC1/HN) of P. falciparum. The PCR product was purified and digested with BamHⅠ, recombinant plasmid pVXORF1-PvMSP 1.19-HBS was digested with the same restriction endonuclease. (2) The plasmid pVXORF1 and recombinant plasmid PET28α-MSP 2 were digested with BamHⅠand XhoⅠ. The two generated DNA fragments in each of the two procedures above were purified and ligated with T4 DNA ligase (according to the rate of MSP 2∶pVXORF1-HBS or MSP 2∶pVXORF1=3∶1) at 16℃ for 16 h, and transferred into Escherichia coli (E. coli) strain DH5α respectively. The recombinant plasmids were screened and identified by BamHⅠand/or XhoⅠand PCR amplification. Results Both the recombinant plasmids pVXORF1-PfMSP 2-HBS and pVXORF1-PfMSP 2 contained the MSP 2 gene from FCC1/HN isolate of P. falciparum. Conclusion The gene encoding MSP 2 was amplified from genomic DNA of recombinant plasmid PET28α-MSP 2; pVXORF1-PfMSP 2-HBS and pVXORF1-PfMSP 2 recombinant were successfully constructed.更多还原