title: Construction and identification of the recombinant lentiviral expression vector targeting human BAX inhibitor-1 gene.
abstract: OBJECTIVE
  The aim of this study was to construct a recombinant lentiviral expression vector targeting human BAX inhibitor- 1(BI-1) gene and observe its expression in NIH3T3 cells.
  
  
  METHODS
  Human BI-1 gene was amplified by polymerase chain reaction (PCR), and then cloned into the vector pLCMV- IG using DNA recombinant technique. After the inserted sequences in the recombinant plasmids were identified by PCR, and double digesting and DNA sequencing analysis, the recombinant lentivirus was packaged and administered into NIH3T3 cells. The BI-1 mRNA and protein expression were examined by real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting.
  
  
  RESULTS
  PCR double digesting analysis and DNA sequencing confirmed that the BI-1 DNA sequences were successfully inserted into the lentiviral vectors. After transfection with the recombinant lentivirus, BI-1 expression in NIH3T3 cells was significantly increased at both mRNA and protein levels.
  
  
  CONCLUSIONS
  The lentiviral vector expressing BI-1 has been successfully constructed, which allowed for the subsequent analysis of the role of BI-1 in cell growth and transduction.
authors.affiliation: Center of Oncology, The Affiliated Hospital of Guangdong Medical College, Guangdong, China.
authors.forename: Hai-qing
authors.initials: HQ
authors.lastname: Luo
authors.forename: Wang-tao
authors.initials: WT
authors.lastname: Zhong
authors.forename: Fu-mei
authors.initials: FM
authors.lastname: Liu
authors.forename: Zhong-yi
authors.initials: ZY
authors.lastname: Cui
authors.forename: Ke-yuan
authors.initials: KY
authors.lastname: Zhou
authors.forename: Xiang-yong
authors.initials: XY
authors.lastname: Li
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular / Q000379:methods; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: D051017:Apoptosis Regulatory Proteins; D008565:Membrane Proteins; D012333:RNA, Messenger; C484209:TMBIM6 protein, human