title: [Construction of adenoviral vectors expressing miR-30a and miR-30e].
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 AdEasy1. 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.
  
  
  CONCLUSIONS
  We have successfully constructed the adenoviral vectors expressing the mature miRNAs.
authors: Clinical Laboratory of the Affiliated Hospital of Jiangsu University, Zhenjiang, China. liuqiang_free@163.com
authors: Qiang
authors: Q
authors: Liu
authors: Jinjin
authors: J
authors: Gu
authors: Min
authors: M
authors: Luo
authors: Qiong
authors: Q
authors: Shi
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D010957:Plasmids
chemical_list: D035683:MicroRNAs; C534715:Mirn30d microRNA, mouse