南方医科大学学报2013，Vol.33Issue(2)：202-206,5.DOI:10.3969/j.issn.1673-4254.2013.02.09
miRNA-30a/30e腺病毒表达载体的构建
Construction of adenoviral vectors expressing miR-30a and miR-30e
摘要
Abstract
Objective To construct adenoviral vectors expressing mature miRNA-30a and miRNA-30e. Methods The target mmu-miR-30a and mmu-miR-30e genes amplified from mouse genome were digested and linked to the shuttle plasmid pSES-HUS, which was then transformed into competent AdEaseier cells for recombination. The confirmed recombinant plasmids were transfected into Hek-293 cells for production of the adenoviruses pAd-mmu-miR-30a and pAd-mmu- miR-30e. The obtained adenoviruses were used to infect Mefs cells, and the cellular expressions of mmu-miR-30a and mmu- miR-30e were detected using fluorescence quantitative PCR. Results mmu-miR-30a (357 bp) and mmu-miR-30e (324 bp) containing the restriction sites were amplified and linked to the shuttle plasmid pSES-HUS, which was successfully recombined with AdEasyl. After packaging in Hek-293 cells, the adenoviral vectors were obtained, which caused an increase of mmu-miR-30a expression by 26.46±7.46 folds and mmu-miR-30e expression by 2.76±0.25 folds in transfected Mefs cells. Conclusion We have successfully constructed the adenoviral vectors expressing the mature miRNAs.关键词
miRNA-30a/miRNA-30e/AdEasy系统/荧光定量PCRKey words
miRNA-30a/miRNA-30e/AdEasy system/fluorescence quantitative polymerase chain reaction引用本文复制引用
刘强,顾金金,罗敏,施琼..miRNA-30a/30e腺病毒表达载体的构建[J].南方医科大学学报,2013,33(2):202-206,5.基金项目
国家自然科学基金(30872770) （30872770）
