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运用OL PCR(OverlapPCR)方法 ,扩增出D2 0 4和D2 MON5 0 1结构蛋白区、5′非编码区等区域的嵌合片段 ,以此片段替换 pDVWS5 0 1质粒中的相应部分后 ,转化DH5α菌。测序结果表明 ,已成功构建含有D2 0 4株完整的PrM和E基因、一部分C基因 ,及D2 MON5 0 1株非编码区、非结构蛋白区和大部分C基因的嵌合病毒全长cD NA(QH 0 4/MON5 0 1)克隆。为进一步研究登革 2型病毒结构蛋白区对毒力的影响打下基础
Using OL PCR (OverlapPCR) method, the chimeric fragments of D2 0 4 and D2 MON 5 0 1 structural protein region and 5 ’non-coding region were amplified and replaced with the corresponding fragment of pDVWS5 0 1 plasmid. DH5α bacteria. The sequencing results showed that the full-length chimeric virus containing D2 0 4 complete PrM and E genes, some C genes and D2 MON5 0 1 non-coding regions, non-structural protein regions and most of the C genes has been successfully constructed (QH 0 4 / MON5 0 1) Clone. This will lay the foundation for further study on the influence of dengue type 2 virus structural protein region on virulence