[Objective] The aims of this study were to explore the expression of KLF15 gene in tissues and differentiation of C2C12 cell of mice and conduct its adenovirus vector:[Method] Firstly,tissues of mice and C2C12 cells in different differentiation phases were collected to extract RNA for detecting expression of KLF15 gene.Secondly,the target fragment obtained from p cDNA3.1-KLF15 vector was connected to pAdTrack-CMV after being digested by Hind Ⅲ and Xba Ⅰ to obtain pAdTrack-KLF15 vector.The pAdTrack-KLF15 vector was linearized by Pme Ⅰ and transformed into BJ5183 competent cells with pAdEasy to construct pAdEasy-KLF15.After the pAdEasy-KLF15 vector was digested by Pac Ⅰ,the purified product was transfected into 293A cells using lip for viral packaging.Finally,q-PCR was used to detect the expression of KLF15 mRNA after pAdEasy-KLF15 vector was injected into C2C12 cells for 48 h.[Result] KLF15 showed significantly increasing expression during differentiation of C2C12 myoblasts and KLF15 expression levels in liver,kidney and fat were significantly higher than in the skeletal muscle and spleen.The recombinant adenovirus vector pAdEasy-KLF15 was successfully obtained,and the mRNA expression of KLF15 gene indicated an observably up-regulated trend after it was transfected C2C12 cells.[Conclusion] KLF15 gene showed high expression in C2C12 cells.The recombinant adenovirus vector expressing mouse KLF15 gene was successfully constructed.
NETL
NSTL
维普
万方数据
