title: [Construction and identification of mouse wildtype-syndecan-1 and unshedding-syndecan-1 expressing vector].
abstract: OBJECTIVE
  To construct the expressing vector pcDNA3.0- wildtype-syndecan-1(WT-sdc1) and pcDNA3.0-unshedding-syndecan-1(uS-sdc1), and to explore the expression in vitro.
  
  
  METHODS
  (1)The mouse WT-sdc1 DNA was successfully amplified by PCR and then cloned into pcDNA3.0. uS-sdc1 was construct by Gene splicing by overlap extension- PCR on the basis of WT-sdc1. The two vectors confirmed by DNA sequencing. (2)there are 4 groups in our research: control group, pcDNA3.0 transfected group, WT-sdc1 transfected group and uS-sdc1 transfected group. each vecter was transfected into IEC-6 cells by Lipofectamine(TM);2000. RT-PCR, Western blot and Dot blot were performed to detect the expression of syndecan-1 before and after stimulation of phorbol 12-myristate 13-acetate (PMA) for 15 min.
  
  
  RESULTS
  The vector WT-sdc1 and uS-sdc1 were successfully constructed although an non-sense mutation was in uS-sdc1. Compared to control and pcDNA3.0 transfected groups, WT-sdc1 and uS-sdc1 groups showed a significant increase in the expression of syndecan-1 in both mRNA and protein levels. In response to the stimulation of PMA, the expression of syndecan-1 was down-regulated at the protein levels but not mRNA levels.
  
  
  CONCLUSIONS
  The WT-sdc1 and uS-sdc1 are successfully constructed, which lays the foundation for further studying of syndecan-1 in gastrointestinal inflammation.
authors.affiliation: Guangdong Provincial Key Laboratory of Gastroenterology,  Department of Gastroenterology,  Nanfang Hospital,  Southern Medical University,  Guangzhou 510515, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.forename: Ji-de
authors.initials: JD
authors.lastname: Wang
authors.forename: Jie-Si
authors.initials: JS
authors.lastname: Kuang
authors.forename: Shao-Heng
authors.initials: SH
authors.lastname: Zhang
authors.forename: Yuan-Lin
authors.initials: YL
authors.lastname: Tang
authors.forename: Zhong-Qiu
authors.initials: ZQ
authors.lastname: Wang
authors.forename: He-Ping
authors.initials: HP
authors.lastname: Qin
authors.forename: Ye
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D015870:Gene Expression / Q000187:drug effects / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053668:Syndecan-1 / Q000235:genetics* / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D053668:Syndecan-1; D013755:Tetradecanoylphorbol Acetate