[Cloning and expression of murine alpha1,2-fucosyltransferase gene].
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 H & Sec1 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.
Lin
Bei
B
Department of Obstetrics and Gynecology, The Second Affiliated Hospital of China Medical University, Shenyang 110004. linbei88@hotmail.com
Saito
Makiko
M
Iwamori
Masao
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D005647:Fucosyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D011817:Rabbits; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction
D005647:Fucosyltransferases