title: [Construction of the eukaryotic expression vector pEGFP-claudin-1 and its expression in 293T cells].
abstract: OBJECTIVE
  To construct the eukaryotic expression vector pEGFP-claudin-1 and to make it express expressed in 293T cells.
  
  
  METHODS
  Claudin-1 ORF was amplified by reverse transcription polymerase chain reaction (RT-PCR). The eukaryotic expression vector pEGFP-claudin-1 was constructed by introducing claudin-1 DNA fragment into the sites of Xho I and BamH I of pEGFP-C3 vector. The plasmid was transfected into 293T cells with lipofectamine. The expressed EGFP was observed by fluorescent microscope. The EGFP-claudin-1 was detected by Western blot.
  
  
  RESULTS
  The eukaryotic expression vector pEGFP-claudin-1 was constructed successfully. The expression of EGFP-claudin-1 was observed in the membrane of transfected cells by fluorescent microscope and a 49 kD protein was detected by Western blot.
  
  
  CONCLUSIONS
  The recombinant expression vector pEGFP-claudin-1 has been successfully constructed and claudin-1 gene can be expressed in 293T cells. This study lays a foundation for further research into the function of claudin-1.
authors.affiliation: Department of Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China. weixinxp@gmail.com
authors.forename: Xin
authors.initials: X
authors.lastname: Wei
authors.forename: Ye
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.forename: Lei
authors.initials: L
authors.lastname: Pan
authors.forename: Ping-zhong
authors.initials: PZ
authors.lastname: Wang
authors.forename: Xue-fan
authors.initials: XF
authors.lastname: Bai
authors.forename: Zhan-sheng
authors.initials: ZS
authors.lastname: Jia
mesh_terms: D000818:Animals; D015153:Blotting, Western; D062445:Claudin-1; D003001:Cloning, Molecular; D006224:Cricetinae; D056890:Eukaryota / Q000235:genetics* / Q000502:physiology; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D005822:Genetic Vectors; D006801:Humans; D008055:Lipids / Q000494:pharmacology; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods; D019444:Vaccines, DNA
chemical_list: C568108:CLDN1 protein, human; D062445:Claudin-1; D008055:Lipids; C086724:Lipofectamine; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D019444:Vaccines, DNA