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Comparison of the Three Murine GDP-fucose: β-galactoside α1，2-fucosytransferase MFUT-Ⅰ， MFUT-Ⅱ and MFUT-Ⅲ
Bei Lin*, Makiko Saito1, Masao Iwamori2
Department of Gynecology and Obstetrics, the Second Affiliated Hospital of China Medical University, Shenyang 110004, China; 1Department of Pediatrics, Graduate School of medicine, University of Tokyo, Tokyo 113-0003, Japan; 2Department of Biochemistry, F
Abstract: Three members of a GDP-fucose: b-galactoside α1，2-fucosyltransferase (α1，2-fucosyltransferase， α1，2-FT)， MFUT-I， MFUT-II， and MFUT-III， from a cDNA of murine small intestine， were cloned by RT-PCR. MFUT-I， MFUT-II， and MFUT-III exhibited sequence homology together and with the human H (78.4%)， Se (79.0%)， and Sec1 (74.9%) gene products， respectively. The open reading frames were ligated into mammalian expression vector pcDNA 3.1(pcDNA3.1-MFUT-I， pcDNA3.1-MFUT-II， pcDNA3.1-MFUT-III，) and then transiently transfected into COS-7 cells using a Cellphect transfection kit and the cells was used for determination of α1，2-FT. COS-7 cells transfected with MFUT-I and MFUT-II exhibited α1，2-FT activity， but no activity was detected in COS-7 cells with MFUT-III. The expression of three gene in murine tissues were analyzed by Northern blotting. MFUT-II yielded a 3.5 kb mRNA transcript in several tissues， whereas MFUT-I and MFUT-III were predominantly expressed in epididymis and testis， respectively. Southern blot analysis showed that MFUT-II was present in the mouse genome as a single-copy gene but MFUT-I and MFUT-III were present as tow-copy gene. Our results suggest that MFUT-I， MFUT-II and MFUT-III correspond to the human H， Se and Sec1 genes， respectively.
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