摘要
目的构建小鼠IL-10启动子荧光素酶报告基因。方法 PCR扩增小鼠IL-10启动子序列,将其插入荧光素酶报告基因pGL3-basic,酶切鉴定及测序比对。结果重组体双酶切电泳后出现大小约4 800 bp与682 bp的两个片段,测序结果显示克隆的小鼠IL-10启动子序列与Genbank中的一致。结论成功构建小鼠IL-10启动子荧光素酶报告基因pGL3-IL10,为进一步研究IL-10奠定了基础。
Objective To construct the luciferase expression vector containing mouse IL-10 promoter. Methods The fragment of IL-10 promoter was amplified from mouse genomic DNA by polymerase chain reaction（PCR）.The amplified fragment was subsequently cloned into PGL3-basic.The recombinant plasmid was confirmed by restriction enzyme digestion and sequencing comparison. Results There were two fragments（4 800 bp and 682 bp） in electrophoresis after restriction enzyme digestion.The result of DNA sequencing showed that the sequence of the cloned IL-10 promoter was identical to that GeneBank had reported. Conclusions The luciferase expression vector containing mouse IL-10 promoter pGL3-IL10 is constructed successfully,and it will become the essential material for further study.
出处
《实用预防医学》
CAS
2011年第12期2252-2253,共2页
Practical Preventive Medicine
基金
国家自然科学基金项目(30672035)
