Analysis of the sequences of human beta-1,4-galactosyltransferase cDNA clones.

S K Chatterjee, S Mukerjee, P K Tripathi

Index: Int. J. Biochem. Cell Biol. 27(3) , 329-36, (1995)

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Abstract

UDP-galactose:beta-1,4 N-acetyl glucosamine galactosyltransferase (4 beta GT) is a promising tumor marker for ovarian cancer. To study the role of 4 beta GT in malignant transformation at the molecular level human 4 beta GT cDNA and genomic clones were isolated and analyzed. For the isolation of 4 beta GT cDNA and genomic clones, a human fetal liver cDNA library in lambda gt11 and a human genomic library in EMBL-3B vectors respectively were screened using a 4 beta GT cDNA insert as the probe. Complete sequence of the cDNA clones were determined by subcloning in plasmid vectors, and compared with the published sequence of human liver 4 beta GT. Presence of various 4 beta GT exons in the genomic clones were determined by Southern blot analysis using specific oligodeoxynucleotide probes. Among the 5 cDNA clones isolated, 2 clones GTN 6 and GTN 17 were sibling clones and had a nucleotide sequence identical to the published 4 beta GT cDNA sequence, except at the 3'-end, where these clones had 7 unique nucleotide sequences. One cDNA clone, GTN2 also had a nucleotide sequence identical to that of 4 beta GT, except for 3 G residues at the 5'-end. One cDNA clone, GTN 1, had a unique sequence at the 5'-end comprising of 74 nucleotides. Another clone, GTN 20, was unrelated to 4 beta GT. Analysis of genomic clones showed that 4 beta GT exons 3, 4, 5 and 6 were present in a 14 kb genomic clone, EMGT-4. Exon 1 was present in a separate 16 kb clone, EMGT-6.(ABSTRACT TRUNCATED AT 250 WORDS)