title: [Construction and effect identification of MiR RNAi eukaryotic expression vectors of prohibitin].
abstract: OBJECTIVE
  To construct the RNAi eukaryotic vector of inhibitory member of the prohibitin (PHB-1) gene and observe the interfering effect in HEK293 cell line after the vector transfection.
  
  
  METHODS
  The specific Mi RNA sequence was designed according to the PHB-1 sequence in GenBank, complementary single-strand DNA oligonucleotides were designed and synthesized, and annealed the single-stranded oligonucleotides to generate a double strands oligonucleotides , cloned the oligonucleotides into pcDNATM6.2-GW/EmGFP-MiR-PHB to obtain an entry clone and then sequence analysis was performed. The recombinant plasmid was transfected into HEK293 cell by liposome. PHB-1 expression was detected by Western blotting.
  
  
  RESULTS
  The DNA sequence of interest clone to the vector was constructed to generate an entry clone and an expression clone successfully, which were proved by sequence determination. Western blotting analysis demonstrated that PHB-1 MiR RNA expression construction could suppress the expression of PHB-1.
  
  
  CONCLUSIONS
  A RNAi eukaryotic vector containing prohibitin gene was successfully constructed.
authors.affiliation: Department of Stress Medicine, Institute of Health & Environmental Medicine, Tianjin, China.
authors.forename: Dong-Sheng
authors.initials: DS
authors.lastname: Guo
authors.forename: Xin-Xing
authors.initials: XX
authors.lastname: Wang
authors.forename: Xiao-Hua
authors.initials: XH
authors.lastname: Liu
authors.forename: Ju-Xiang
authors.initials: JX
authors.lastname: Yuan
authors.forename: Ling-Jia
authors.initials: LJ
authors.lastname: Qian
mesh_terms: D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics; D000091402:Prohibitins; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
chemical_list: D035683:MicroRNAs; C000717328:PHB protein, human; D000091402:Prohibitins; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D012097:Repressor Proteins