Molecular cloning, expression and exon/intron organization of the bovine β-galactoside α2,6-sialyltransferase gene

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作者：

Dominique，Mercier，Anne，Wierinckx，Ahmad，Oulmouden，Paul，F.，Gallet，Monica

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摘要：

In this study, we report the first isolation and characterization of a bovine sialyltransferase gene. Bovine cDNAs prepared from different tissues contain an open-reading frame encoding a 405 amino acid sequence showing 83%, 75%, and 60% identity with human, murine, and chicken ST6Gal I (β-galactoside α2,6-sialyltransferase) sequences, respectively. When transfected into COS-7 cells, a recombinant enzyme was obtained which catalyzed the in vitro α2,6-sialylation of Lac-NAc (NeuAcα2-6Galβ1–4GlcNAc) and LacdiNAc (NeuAc-α2-6GalNAcβ1-4GlcNAc) acceptor substrates. The Km values were 2.8 and 6.9 mM, respectively. Different relative efficiencies (V<sub>max</sub>/K<sub>m</sub>) for the two precursors (36 for LacNAc and 4.3 for LacdiNAc) were observed. Bovine ST6Gal I gene consists of four 5′-untranslated exons E(−2) to E(1), and five coding exons from E(2) to E(6). This later carries a 3′-untranslated region of 2.7 kb. Gene sequence spans at least 80 kb of genomic DNA. Two processed pseudogenes have been identified. They are 94.3 and 95.6% similar to the bovine cDNA, respectively. Three families of mRNA isoforms were isolated. They differed by their 5′-untranslated regions and could be generated by three tissue-specific promoters. Family 1 is made up of exons E(−2) and E(1) to E(6), family 2 of exons E(−1) to E(6), and family 3 of exons E(1) to E(6). Tissular distribution of transcript families appears noticeably different than those described in human and rat.

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关键词：

bovine sialyltransferase ST6Gal I italic&gt gene LacNAc LacdiNAc