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Molecular cloning and expression of chick Gal beta 1,3GalNAc alpha 2,3-sialyltransferase
N Kurosawa1, T Hamamoto, M Inoue
1Frontier Research Program, Institute of Physical and Chemical Research (RIKEN), Saitama, Japan.
Abstract:
A cDNA clone encoding chick Gal beta 1,3GalNAc alpha 2,3-sialyltransferase (ST3Gal I) was isolated from a chick embryo brain cDNA library. The cDNA sequence included an open reading frame coding for 342 amino acids, and the deduced amino acid sequence showed 64% identity with that of the mouse enzyme. Northern blot analysis of chick embryos revealed that the ST3Gal I gene was expressed in early embryonic stages. The identity of the enzyme was confirmed by construction of a recombinant sialyltransferase in which the N-terminal part including the cytoplasmic tail and signal anchor domain was replaced with an immunoglobulin signal peptide sequence. This enzyme expressed in COS-7 cells exhibited transferase activity similar to that of mouse ST3Gal I.