title: Cloning and expression of mouse peroxiredoxin I in IEC-6 Cells.
abstract: OBJECTIVE
  To clone and express mouse peroxiredoxin I in IEC-6 cells.
  
  
  METHODS
  Total RNAs were isolated from cultured IEC-6 cells, and the coding region of peroxiredoxin I was amplified by RT-PCR. After it was cloned into T-vector and sequenced, pSG5 was used to transiently express peroxiredoxin I in IEC-6 by liposome-mediated transfection, and the expression of peroxiredoxin I was evaluated by RT-PCR and Western blot.
  
  
  RESULTS
  A DNA fragment about 750 bp was amplified from total RNAs of IEC-6 cells using specific primers of peroxiredoxin I. The sequencing confirmed the coding region was successfully cloned into T-vector, which was completely coincident with the sequence in GeneBank. After the EcoRI-BamHI fragment of T-vector containing peroxiredoxin I was inserted into pSG5, the recombinant plasmid was transferred to IEC-6 cells. RT-PCR assay showed that a DNA fragment of 930 bp could be amplified, which indicated the transcription of pSG5-Prx. Western blot confirmed the expression of peroxiredoxin I in IEC-6 cells.
  
  
  CONCLUSIONS
  Mouse peroxiredoxin I can be successfully expressed in IEC-6 cells.
authors.affiliation: Institute of Combined Injury of PLA, State Key Laboratory of Trauma, Third Military Medical University, Gaotanyan Street 30, Chongqing 400038, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.forename: Yong-Ping
authors.initials: YP
authors.lastname: Su
authors.forename: Tao
authors.initials: T
authors.lastname: Wang
authors.forename: Feng-Chao
authors.initials: FC
authors.lastname: Wang
authors.forename: Guo-Ping
authors.initials: GP
authors.lastname: Ai
authors.forename: Hui
authors.initials: H
authors.lastname: Xu
authors.forename: Jun-Ping
authors.initials: JP
authors.lastname: Wang
authors.forename: Yue-Sheng
authors.initials: YS
authors.lastname: Huang
authors.forename: Jian-Xin
authors.initials: JX
authors.lastname: Jiang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary; D015870:Gene Expression; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010544:Peroxidases / Q000235:genetics* / Q000378:metabolism*; D054464:Peroxiredoxins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
chemical_list: D018076:DNA, Complementary; D010544:Peroxidases; D054464:Peroxiredoxins