新型抗炎因子IL-37真核表达载体pIRES2-EGFP/IL-37的构建及鉴定

Construction and identification of the eukaryotic expression vector of IL-37(pIRES2-EGFP/IL-37)

目的构建新型抗炎因子IL-37的真核表达载体p IRES2-EGFP/IL-37(p IEIL37)并进行鉴定。方法 Trizol法提取THP-1细胞RNA,RT-PCR扩增IL-37基因,将目的基因片段定向克隆到真核表达载体p IRES2-EGFP;将重组质粒转化大肠杆菌DH5,并进行PCR、双酶切和DNA测序鉴定。结果琼脂糖凝胶电泳显示1条约710 bp大小的条带;双酶切结果显示约675 bp大小的条带;DNA测序表明IL-37基因正确插入真核表达载体p IRES2-EGFP。结论成功构建IL-37真核表达载体p IRES2-EGFP/IL-37。

Objective To construct and identify the eukaryotic expression vector of IL-37 （pIRES2-EGFP/IL-37）. Methods RNA was extracted from THP-1 cells and IL-37 gene was amplified by RT-PCR. The target gene was cloned into the eukaryotic expression vector plRES2-EGFP. The recombinant plasmid was transfected into Dh5ct strain of Escheria coil The plEIL37 was identified by PCR, double enzyme digestion and DNA sequencing. Results Agarose electrophoresis revealed a 710 bp band. Double enzyme digestion showed a 675 bp band. DNA sequencing indicated the insertion of IL-37 gene into eukaryotic expression vector plRES2-EGFP. Conclusion The eukaryotic expression vector of IL-37 （plRES2-EGFP/IL-37） is successfully constructed.