TY - JOUR
T1 - Molecular cloning and chromosomal localization of human 4-β-galactosyltransferase
AU - Humphreys-Beher, M. G.
AU - Bunnell, B.
AU - VanTuinen, P.
AU - Ledbetter, D. H.
AU - Kidd, V. J.
PY - 1986
Y1 - 1986
N2 - A cDNA clone to human 4-β-galactosyltransferase (EC 2.4.1.38) was isolated from a human liver λgt11 expression library by using a monospecific polyclonal antiserum to affinity-purified bovine enzyme. The authenticity of this cDNA clone has been demonstrated by several criteria. Under conditions of chronic treatment with the β-adrenergic receptor agonist isoproterenol, rat parotid glands show an approximately 10-fold increase in 4-β-galactosyltransferase activity. The increased enzyme activity was reflected in dot-blot analysis of control and isoproterenol-treated rat parotid RNA by using the human cDNA as probe. Hybrid-selection and in vitro translation identified a protein product with a molecular mass of 47 kDa that was immunoprecipitated with the bovine antiserum. The full-length human cDNA clone was then isolated and the DNA sequence for the NH2-terminal portion of the protein was deduced. Comparison of the NH2-terminal protein sequence from the bovine protein with that of the human cDNA clone confirmed its identity. In addition, the human cDNA clone was used to localize the gene for 4-β-galactosyltransferase to human chromosome 4 by Southern analysis of a somatic cell hybrid panel.
AB - A cDNA clone to human 4-β-galactosyltransferase (EC 2.4.1.38) was isolated from a human liver λgt11 expression library by using a monospecific polyclonal antiserum to affinity-purified bovine enzyme. The authenticity of this cDNA clone has been demonstrated by several criteria. Under conditions of chronic treatment with the β-adrenergic receptor agonist isoproterenol, rat parotid glands show an approximately 10-fold increase in 4-β-galactosyltransferase activity. The increased enzyme activity was reflected in dot-blot analysis of control and isoproterenol-treated rat parotid RNA by using the human cDNA as probe. Hybrid-selection and in vitro translation identified a protein product with a molecular mass of 47 kDa that was immunoprecipitated with the bovine antiserum. The full-length human cDNA clone was then isolated and the DNA sequence for the NH2-terminal portion of the protein was deduced. Comparison of the NH2-terminal protein sequence from the bovine protein with that of the human cDNA clone confirmed its identity. In addition, the human cDNA clone was used to localize the gene for 4-β-galactosyltransferase to human chromosome 4 by Southern analysis of a somatic cell hybrid panel.
UR - http://www.scopus.com/inward/record.url?scp=0022908289&partnerID=8YFLogxK
U2 - 10.1073/pnas.83.23.8918
DO - 10.1073/pnas.83.23.8918
M3 - Article
C2 - 3097639
AN - SCOPUS:0022908289
SN - 0027-8424
VL - 83
SP - 8918
EP - 8922
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 23
ER -