351. [Cloning and expression of murine alpha1,2-fucosyltransferase gene]
- Author
-
Bei, Lin, Makiko, Saito, and Masao, Iwamori
- Subjects
Base Sequence ,Reverse Transcriptase Polymerase Chain Reaction ,Molecular Sequence Data ,Blotting, Northern ,Fucosyltransferases ,Gene Expression Regulation, Enzymologic ,Rats ,Blotting, Southern ,Mice ,Open Reading Frames ,COS Cells ,Chlorocebus aethiops ,Animals ,Humans ,Amino Acid Sequence ,Rabbits ,Cloning, Molecular - Abstract
MFUT-II, a DNA fragment encoding murine alpha1,2-fucosyltransferase,was cloned by RT-PCR. The open-reading frame was ligated into mammalian expression vector pcDNA 3.1 (pcDNA3.1-MFUT-II) and was transiently transfected into COS-7 cells using a Cellphect Transfection kit. Moreover, the expression of the gene in murine tissues was analyzed with Northern hybridization showing a 3.5-Kb mRNA transcript product in several tissues. The existence of MFUT-II was detected by Southern blot. MFUT-II was found to be a new member of the murine alpha1,2-FT gene family. Its open reading frame encoded 347 amino acids with a predicted molecular mass of 39.21 kDa and exhibited sequence homology with murine HSec1 gene. MFUT-II also exhibited sequence homology with Human Se gene (79.0%), Rat FTB gene (89.0%) and Rabbit FT-III gene (77.0%), respectively. COS-7 cells transfected with pcDNA3.1-MFUT- II showed alpha 1,2-fucosyltransferase activity. Southern analysis revealed that MFUT-II was present in the mouse genome as a single-copy gene. In conclusion, MFUT-II was a murine Se gene.
- Published
- 2006