title: [Effect of α -1,2 fucosyltransferase gene 682A> G and 547_552delAG mutations on the activity of fucosyltransferase].
abstract: OBJECTIVE
  To explore the effect of α -1,2 fucosyltransferase (FUT1) gene 682A> G and 547_552delAG mutations on the expression of FUT1 mRNA and activity of α -1,2 fucosyltransferase.
  
  
  METHODS
  Recombinant expression vectors of FUT1 682A> G and FUT1 547_552delAG were constructed and transfected into COS-7 cells for stable expression screening. Expression of FUT1 mRNA was determined using real-time quantitative PCR. The activity of FUT1 was measured with high-performance liquid chromatography.
  
  
  RESULTS
  Stably transfected COS-7 cells with wild type FUT1, FUT1 682A> G and FUT1 547_552delAG were respectively obtained. The FUT1 mRNA level of transfected cells with 682A> G and 547_552delAG recombination vectors have measured 101.69% and 102.79% compared with that of wild type FUT1 transfected cells. A specific protein band with about 46 kD was confirmed in the 682A> G transfected cell lysates by SDS-PAGE electrophoresis and Western blotting with 6× His Tag antibody. Similar protein was not identified in the 547_552delAG cells lysates. Enzymes activity of FUT1 682A> G has measured 61.01% compared with wild type FUT1 protein, whilst the activity of FUT1 547_557delAG was completely abolished.
  
  
  CONCLUSIONS
  FUT1 682A> G and 547_552delAG mutations do not affect the transcript efficiency, although various mutations have different impact on the enzyme's activity.
authors.affiliation: Institute of Transfusion Medicine, Blood Center of Zhejiang Province, Key Laboratory of Blood Safety Research, Ministry of Health, Hangzhou, Zhejiang 310006, P.R.China. zfm00@hotmail.com.
authors.forename: Sudan
authors.initials: S
authors.lastname: Tao
authors.forename: Yanmin
authors.initials: Y
authors.lastname: He
authors.forename: Xianguo
authors.initials: X
authors.lastname: Xu
authors.forename: Xiaozhen
authors.initials: X
authors.lastname: Hong
authors.forename: Ji
authors.initials: J
authors.lastname: He
authors.forename: Faming
authors.initials: F
authors.lastname: Zhu
authors.forename: Hangjun
authors.initials: H
authors.lastname: Lyu
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D019556:COS Cells; D002522:Chlorocebus aethiops; D002851:Chromatography, High Pressure Liquid; D004252:DNA Mutational Analysis; D005647:Fucosyltransferases / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D009154:Mutation; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000097631:Galactoside 2-alpha-L-fucosyltransferase
chemical_list: D005647:Fucosyltransferases; D011994:Recombinant Proteins; D000097631:Galactoside 2-alpha-L-fucosyltransferase