| 摘要 |
A novel gene defining a novel enzyme in the UDP-D-galactose: b-N-acetyl-glucosamine beta-1,4-galactosyltransferase family, termed beta4Gal-T2, with unique enzymatic properties is disclosed. The enzymatic activity of beta4Gal-T2 is shown to be distinct from that of previously identified enzymes of this gene family. The invention discloses isolated DNA molecules and DNA constructs encoding beta4Gal-T2 and derivatives thereof by way of amino acid deletion, substitution or insertion exhibiting beta4Gal-T2 activity, as well as cloning and expression vectors including such DNA, cells transfected with the vectors, and recombinant methods for providing beta4Gal-T2. The enzyme beta4Gal-T2 and beta4Gal-T2-active derivatives thereof are disclosed, in particular soluble derivatives comprising the catalytically active domain of beta4Gal-T2. Further, the invention discloses methods of obtaining beta-1,4-galactosyl glycosylated saccharides, glycopeptides or glycoproteins by use of an enzymically active beta4Gal-T2 protein or fusion protein thereof or by using cells stably transfected with a vector including DNA encoding an enzymatically active beta4Gal-T2 protein as an expression system for recombinant production of such glycopeptides or glycoproteins. Also a method for the identification of DNA sequence variations in the beta4Gal-T2 gene by isolating DNA from a patient, amplifying beta4Gal-T2-coding exons by PCR, and detecting the presence of DNA sequence variation, are disclosed.
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