棉铃虫核多角体病毒进化保守性基因iap2基因表达载体的构建及表达

分析棉铃虫核多角体病毒基因组 ,结合GenBank中已知的序列 ,发现iap2基因位于其基因组的BamHⅠ F片段上 ,回收此片段作为模板 ,设计引物 ,通过PCR扩增得到了抗细胞凋亡基因iap2的DNA片段。将扩增产物克隆到pGEM T载体上 ,再进一步将插入片段酶切并连接到表达载体pET 2 8a上 ,构建了重组质粒pET iap2。DNA序列分析结果表明 ,克隆得到的DNA序列与所发表序列完全相同。含重组质粒pET iap2的大肠杆菌BL2 1 (DE3)表达了抗细胞凋亡蛋白IAP2。

Coloning HaSNPV iap2 Gene and its expression in prokaryotic expression system

The iap2 gene of HaSNPV was located in the BamHⅠ-F fragment trough. The genome of HaSNPV and combined this information with the known sequences already available from GenBank was analyzed. Then the BamHⅠ-F fragment as a template and design primer to obtain the DNA fragment of the iap2 gene by PCR amplification were used. Firstly, the blunt-end PCR results were ligated into a pGEM-T vector, then the fragment was transformed into the expression vector pET-28a and digested with BamHⅠand EcoRⅠ. The results of DNA sequence analysis demonstrated that the obtained DNA sequence was the same with the reported one. IAP2 protein was expressed in BL21, which has the recombinant plasmid, pET-iap2.