title: [Cloning of IPO8 promoter and analysis of its transcription activity].
abstract: OBJECTIVE
  To clone 5' untranslated region of human IPO8 gene and determine its transcription activity.
  
  
  METHODS
  We used 5' rapid amplification of cDNA ends (RACE) analysis to identify the IPO8 transcription start site (TSS), and amplified series truncated 5' UTR fragment containing transcription start site. The PCR productions were inserted into luciferase report vector pGL3- Basic. After confirmation by restriction enzyme digestion, the recombinant plasmids were cotransfected into Saos-2 cells with plasmid pRL-TK. The luciferase activities were measured by dual luciferase reporter system.
  
  
  RESULTS
  The IPO8 gene transcription start site was established. The electrophoresis analysis of restriction enzyme digestion and DNA sequencing verified the fragments were successfully amplified and inserted into pGL3-Basic. After the recombinant plasmids transfected, the highexpressions of luciferase were detected in Saos-2 cells.
  
  
  CONCLUSIONS
  The recombinant vector containing IPO8 promoter is constructed successfully, which provides a foundation for determining expressional regulation of IPO8 in the further study.
authors.affiliation: Department of Pharmacology, College of Basic Medical Science, Jiujiang University, Jiujiang Jiangxi 332000, China.
authors.forename: Jianjun
authors.initials: J
authors.lastname: Xiong
authors.forename: Zhen
authors.initials: Z
authors.lastname: Gong
authors.forename: Xiao'ou
authors.initials: X
authors.lastname: Zhou
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.forename: Jianyun
authors.initials: J
authors.lastname: Liu
authors.forename: Weidong
authors.initials: W
authors.lastname: Li
mesh_terms: D003001:Cloning, Molecular; D018076:DNA, Complementary; D005822:Genetic Vectors; D006801:Humans; D008156:Luciferases; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014162:Transfection; D028961:beta Karyopherins / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; C107282:IPO8 protein, human; D028961:beta Karyopherins; D008156:Luciferases