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Volume 270, Number 15, Issue of April 14, pp. 8844-8850, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Molecular Cloning and Expression of Two Types of Rabbit -Galactoside 1,2-Fucosyltransferase

Seiji Hitoshi , Susumu Kusunoki , Ichiro Kanazawa , Shuichi Tsuji

Two DNA clones encoding rabbit -galactoside 1,2-fucosyltransferase (RFT-I and RFT-II) have been isolated from a rabbit genomic DNA library. The DNA sequences revealed open reading frames coding for 373 (RFT-I) and 354 (RFT-II) amino acids, respectively. The deduced amino acid sequences of RFT-I and RFT-II showed 56% identity with each other, and that of RFT-I showed 80% identity with that of human H blood type 1,2-fucosyltransferase. Northern blot analysis of embryo and adult rabbit tissues revealed that the RFT-I gene was expressed in adult brain, and that the RFT-II gene was expressed in salivary and lactating mammary glands. The identities of these enzymes were confirmed by constructing recombinant fucosyltransferases in which the N-terminal part including the cytoplasmic tail and signal anchor domain was replaced with the immunoglobulin signal peptide sequence. RFT-I expressed in COS-7 cells exhibited similar transferase activity to that of human H blood type 1,2-fucosyltransferase. RFT-II expressed in COS-7 cells showed higher affinity for type 1 (Gal1,3GlcNAc) and type 3 (Gal1,3GalNAc) acceptors than type 2 (Gal1,4GlcNAc) ones, which suggested that RFT-II was a putative secretor-type 1,2-fucosyltransferase.




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