Analysis of the sequences of human beta-1,4-galactosyltransferase cDNA clones.
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)
Chatterjee
S K
SK
Department of Obstetrics and Gynecology, University of Kentucky, Lexington 40536-0096, USA.
Mukerjee
S
S
Tripathi
P K
PK
D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000235:genetics; D015139:Blotting, Southern; D003001:Cloning, Molecular; D015342:DNA Probes; D004262:DNA Restriction Enzymes / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D000939:Epitopes / Q000276:immunology; D005091:Exons; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D009237:N-Acetyllactosamine Synthase / Q000737:chemistry / Q000235:genetics*; D016366:Open Reading Frames; D010051:Ovarian Neoplasms / Q000235:genetics; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D014408:Biomarkers, Tumor; D015342:DNA Probes; D018076:DNA, Complementary; D000939:Epitopes; D007074:Immunoglobulin G; D009237:N-Acetyllactosamine Synthase; D004262:DNA Restriction Enzymes