2010•Zhongguo xiandai yixue/Zhongguo xiandai yixue zazhiRequires access
Construction and identification of luciferase expression vector containing mouse TNF-α promoter
Cao Hai-mei
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Abstract
[Objective] To construct the luciferase expression vector containing mouse TNF-α promoter. [Methods]The fragment of TNF-α promoter was amplified from mouse genomic DNA by Polymerase Chain Reaction (PCR). The amplified fragment was subsequently cloned into PGL3-basic. The recombinant pGL3-TNFα-promoter was transfected into 264.7 cells by lipofectamine after it was confirmed. The activity of luciferase was detected.[Results]The result of DNA sequencing showed that the sequence of the cloned TNF-α promoter was identical to what GeneBank reported. The luciferase activity demonstrated that the PGL3-TNF-α promoter had the promoter activity. [Conclusion]The luciferase reporter gene containing mouse TNF-α promoter pGL3-TNF-α promoter is constructed successfully, and it will become essential material for further study.