Isolation, Characterization, and Mapping of a Human Acid β-Galactosidase cDNA

Yoshimi Yamamoto, Cynthia A. Hake, Brian M. Martin, Keith A. Kretz, Amelia J. Ahern-Rindell, Susan L. Naylor, Michal Mudd, John S. O'brien

Research output: Contribution to journalArticlepeer-review

50 Scopus citations

Abstract

A λgt11 human testicular cDNA library was screened with degenerate oligonucleotide probe mixtures based on amino acid sequence data generated from cyanogen bromide fragments and tryptic fragments of purified human β-galactosidase. Six positive clones were identified after screening 2 × 106 plaques. The sequences of these six clones were determined and found to be derived from two different cDNAs. The sequence of the longest of these cDNAs is nearly identical to that recently determined by Oshima et al. (1988). It codes for a 76-kD protein and all 11 peptides that were generated from the purified enzyme. The second clone is shorter by 393 bp in the central portion of the coding region. Analysis by Northern blotting revealed the presence of a single mRNA species of 2.45 kb in lymphoblasts and testicular tissue. It is deduced from the amino acid sequence data that proteolytic processing of the precursor form of β-galactosidase must occur by cleavage in the carboxy-terminal portion of the polypeptide perhaps around amino acid 530 at a uniquely hydrophilic sequence. Using a probe generated from the 3′ region of the cDNA, we have mapped the locus coding for human β-galactosidase to chromosome 3p21-3pter.

Original languageEnglish (US)
Pages (from-to)119-127
Number of pages9
JournalDNA and Cell Biology
Volume9
Issue number2
DOIs
StatePublished - Mar 1990

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

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