摘要
目的 构建恶性疟原虫 FCC- 1/ HN株 MSA2编码基因分枝杆菌穿梭质粒重组子 p BCG5 .6 / MSA2 ,并进行序列测定。 方法 根据恶性疟原虫 MSA2编码基因的两侧序列设计引物 ,采用 PCR技术扩增出 MSA2编码基因全长片段 ,经低熔点琼脂糖挖块法回收纯化后 ,插入分枝杆菌穿梭质粒 p BCG5 .6 / MSA2 ,并转化大肠杆菌 DH5 α,快速酚法初筛阳性重组子 ,阳性克隆以 PCR法与限制性酶切分析鉴定后 ,双脱氧链终止法双向进行序列测定。 结果 从恶性疟原虫基因组中扩增出约 82 0 bp的基因片段 ,所构建 p BCG5 .6 / MSA2重组体阳性克隆经双酶切和 PCR鉴定与预期结果一致 ,测序结果确证了插入片段的正确性。 结论 成功构建了恶性疟原虫 MSA2编码基因分枝杆菌穿梭表达质粒 ,为恶性疟 BCG疫苗的研制奠定了基础。
Objective Construction and sequence determination of recombinant E.coli Mycobacteria shuttle plasmid of MSA2 coding gene of Plasmodium falciparum . Methods The MSA2 gene fragment was amplified by PCR. After purification, the gene fragment was ligated with E.coli Mycobacteria shuttle plasmid pBCG5.6 at polylinker. The recombinant plasmid pBCG5.6/MSA2 was transferred into E.coli DH5α. Positive clones were screened and identified by PCR method and digested with restriction enzyme. The sequence of the inserted MSA2 gene fragment was also determined. Results and Conclusion The recombinant shuttle plasmid of MSA2 gene fragment of P. falciparum was successfully constructed. That would make it convenient for the further research of BCG malaria vaccine.
出处
《中国寄生虫病防治杂志》
CSCD
2002年第1期18-20,共3页
Chinese Journal of Parasitic Disease Control
关键词
恶性疟原虫
裂殖子表面抗原-2编码基因
克隆
序列分析
Plasmodium falciparum
merozoite surface antigen 2 coding gene
DNA clone
sequence determination
