Bovine galactosyltransferase: identification of a clone by direct immunological screening of a cDNA expression library.
A 1.3-kilobase cDNA clone (7A) coding for bovine galactosyltransferase (glycoprotein 4-beta-galactosyltransferase, EC 2.4.1.38) was isolated from a lambda gt11 expression library by immunological screening with monospecific polyclonal antisera to the affinity-purified bovine enzyme. The nucleotide sequence of this clone predicts an open reading frame that starts at the 5' end of the insert and codes for a polypeptide of 334 amino acids with Mr 37,645. Based on a Mr of 57,000 for the membrane-bound enzyme this clone accounts for approximately 61% of the coding sequence. Portions of the predicted amino acid sequence matched the six tryptic peptides isolated from affinity-purified bovine galactosyltransferase. Clone 7A hybridizes to a 4.8-kilobase bovine mRNA and identifies multiple EcoRI restriction fragments in bovine, murine, and human DNA.
N L
NL
Shaper
J H
JH
Shaper
J L
JL
Meuth
J L
JL
Fox
H
H
Chang
I R
IR
Kirsch
G F
GF
Hollis
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D002846:Chromatography, Affinity; D004247:DNA / Q000235:genetics; D004274:DNA, Recombinant / Q000032:analysis; D005700:Galactosyltransferases / Q000235:genetics*; D005819:Genetic Markers; D006801:Humans; D007668:Kidney; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology; D051379:Mice; D012313:RNA / Q000032:analysis; D012689:Sequence Homology, Nucleic Acid; D006022:beta-N-Acetylglucosaminylglycopeptide beta-1,4-Galactosyltransferase / Q000235:genetics* / Q000276:immunology
D000906:Antibodies; D004274:DNA, Recombinant; D005819:Genetic Markers; D008565:Membrane Proteins; D012313:RNA; D004247:DNA; D005700:Galactosyltransferases; D006022:beta-N-Acetylglucosaminylglycopeptide beta-1,4-Galactosyltransferase