Effect of α -1,2 fucosyltransferase gene 682A> G and 547_552delAG mutations on the activity of fucosyltransferase.
- Author:
Sudan TAO
1
;
Yanmin HE
;
Xianguo XU
;
Xiaozhen HONG
;
Ji HE
;
Faming ZHU
;
Hangjun LYU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Base Sequence; Blotting, Western; COS Cells; Cercopithecus aethiops; Chromatography, High Pressure Liquid; DNA Mutational Analysis; Fucosyltransferases; genetics; metabolism; Humans; Mutation; Recombinant Proteins; metabolism; Reverse Transcriptase Polymerase Chain Reaction
- From: Chinese Journal of Medical Genetics 2014;31(5):591-594
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSRecombinant 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.
RESULTSStably 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.
CONCLUSIONFUT1 682A> G and 547_552delAG mutations do not affect the transcript efficiency, although various mutations have different impact on the enzyme's activity.
