IL-37b基因克隆及其真核的表达载体构建

Cloning of region gene of IL-37b and its eukaryotic expression vector construction

目的克隆IL-37b编码区基因,并构建其真核表达载体。方法通过RT-PCR扩增IL-37b编码区基因,DNA测序鉴定后,将阳性克隆提取质粒,通过酶切连接将目的片段定向克隆至真核表达载体pcDNA 3.1,通过菌落PCR和DNA测序等进行鉴定。结果凝胶电泳显示RT-PCR扩增出1条大小约650 bp特异条带,DNA测序结果显示获取了大小为654 bp的IL-37b编码区基因。PCR和DNA测序对所构建的真核表达载体鉴定结果表明IL-37b编码区基因正确插入真核表达载体pcDNA3.1。结论成功克隆了IL-37b编码区基因,并成功构建其真核表达载体,为下一步IL-37b生物学作用的深入研究奠定了基础。

Objective To clone the coding region gene of IL-37b and to construct the eukaryotic expression vector of IL-37b.Methods The coding region gene of IL-37b was amplified by RT-PCR,and the plasmid of positive clone was isolated after identification by DNA sequencing.The target gene was sub-cloned into the eukaryotic expression vector pcDNA3.1,which was identified by PCR and DNA sequencing.Results A specific DNA band about 650 bp of RT-PCR product was seen by agarose electrophoresis,and DNA sequencing result showed that IL-37b gene of 654 bp was achieved.After construction of the eukaryotic expression vector was identified by PCR and DNA sequencing,the result showed that coding region gene of IL-37b was inserted into the eukaryotic vector pcDNA3.1.Conclusion The coding region gene of IL-37b is successfully cloned as well as its eukaryotic expression vector,which lays a foundation for further research of the biology role of IL-37b.