摘要
目的构建真核表达IL-23p193'UTR质粒(在PGL3中),并且进行鉴定,为调查鼠IL-23p193'UTR区对IL-23p19的表达调控提供条件。方法用mp193'UTR/PGL3-controlvectorDNA作为模板,设计不同长度3'UTR的引物,行聚合酶链反应扩增目的基因,将PCR扩增产物回收,酶切,并克隆到PGL3control载体中,通过酶切鉴定重组质粒。结果构建了4个不同长度的小鼠IL-23p193'UTR区的质粒,经酶切测序鉴定,证实与原设计相同。结论成功构建4个不同长度的小鼠IL-23p193'UTR区的质粒,这4个不同长度的质粒含有不同的ARE区,为进一步调查不同ARE区对小鼠IL-23p19的调节作用提供了物质条件。
Objective To construct and identify eukaryotic expression plasmid IL-23p19 3＇UTR（in PGL3） in order to provide conditions for the investigation of the regulation of IL-23p19 by IL-23p193＇UTR in mice.Methods Using mp19 3＇UTR/PGL3 control vector DNA as a template,designing primers of different length of 3＇UTR,polymerase chain reaction amplification of the gene,then the amplification products of PCR were recycled and digested with enzyme,cloned into PGL3 control vector,the recombinant plasmid was identified by restriction enzyme.Results The four different length of IL-23p19 3＇UTR region of plasmids in mice were constructed,which were confirmed the same as the original design by sequencing.Conclusion The four different length of IL-23p19 3 ＇UTR region plasmids in mice are successfully constructed,which contain different ARE areas,so as to provide material conditions for further investigation on the regulatory effect of different ARE on mIL-23p19.
出处
《河北医药》
CAS
2013年第9期1288-1291,共4页
Hebei Medical Journal
基金
河北省应用基础研究计划项目(编号:10966119D)
