title: [Construction of Cyr61 siRNA expression vector and its characteristic investigation].
abstract: OBJECTIVE
  To construct siRNA recombinant expression vector targeting Cyr61 gene, and to screen the stably transfected cell clone.
  
  
  METHODS
  Three siRNAs sequences based on the sequence of Cyr61mRNA in the GenBank were designed and synthesized, and one scrambled siRNA sequences served as negative control. The cDNA was synthesized and inserted into plasmid pGenesil1.1. The plasmids was sequenced to confirm the inserted sequence.RT-PCR and Western blot analysis was used to assess the levels of Cyr61 mRNA and proteins after the constructed plasmids have been transfected into A549 cells.
  
  
  RESULTS
  It was confirmed by restriction endonuclease and sequence analysis that siRNA recombinant expression vector targeting Cyr61 gene was constructed successfully. Inhibition ratio of Cyr61 siRNA at mRNA and protein levels were 59.2% and 52.97%.
  
  
  CONCLUSIONS
  The siRNA recombinant expression vector targeting Cyr61 gene has been constructed successfully and can inhibite the expression of Cyr61 gene in A549 cells significantly.
authors.affiliation: Department of Anesthesiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China. yanzizhang917@126.com
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Qing-ping
authors.initials: QP
authors.lastname: Wu
authors.forename: Shang-long
authors.initials: SL
authors.lastname: Yao
mesh_terms: D015153:Blotting, Western; D055515:Cysteine-Rich Protein 61 / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000502:physiology; D005822:Genetic Vectors; D006801:Humans; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
chemical_list: C501848:CCN1 protein, human; D055515:Cysteine-Rich Protein 61; D034741:RNA, Small Interfering