title: [Construction and expression of the eukaryotic expression vector containing AIF gene].
abstract: OBJECTIVE
  To construct the eukaryotic express vector containing apoptosis-inducing factor (AIF) gene and to study its expression in A549 cells.
  
  
  METHODS
  According to the GenBank AIF mRNA sequence, specific primers to amplify AIF gene from lung carcinoma cell line A549 by RT-PCR was designed. The amplified AIF gene fragment was cloned into plasmid pUC-T by TA cloning, then double enzyme digestion and DNA sequencing were used to identifying the positive recombinant AIF-pUC-T. The target fragment was retrieved and cloned into the eukaryotic express vector pcDNA3.1(+). The positive recombinant AIF-pcDNA3.1(+) was transfected into A549 cells, and expression of AIF gene was verified by RT-PCR and Western blot.
  
  
  RESULTS
  AIF target gene was successfully amplified and cloned into the pUC-T. The target fragment was retrieved and cloned into the eukaryotic express vector pcDNA3.1(+), and it was completely coincided with the AIF sequence in GenBank suggested by cells transfected with AIF-pcDNA3. 1(+) was much higher than that of control cells which was not transfected with AIF-pcDNA3.1(+).
  
  
  CONCLUSIONS
  The AIF eukaryotic expression vector AIF-pcDNA3.1(+) is successfully constructed in A549 cells and it could be experimental foundations for further study of AIF gene.
authors.affiliation: Department of Environmental Health, West China School of Public Health, Sichuan University, Chengdu 610041, China.
authors.forename: Wen-Wen
authors.initials: WW
authors.lastname: Deng
authors.forename: Shu-Kun
authors.initials: SK
authors.lastname: Liu
authors.forename: Zun-Zhen
authors.initials: ZZ
authors.lastname: Zhang
mesh_terms: D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology; D051033:Apoptosis Inducing Factor / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism / Q000473:pathology; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: C494829:AIFM1 protein, human; D051033:Apoptosis Inducing Factor; D012333:RNA, Messenger; D011994:Recombinant Proteins