Isolation of cDNA clones for human β-glucocerebrosidase using the λgtll expression system

Edward I. Ginns, Prabhakara V Choudary, Brian M. Martin, Suzanne Winfield, Barbara Stubblefield, June Mayor, Denise Merkle-Lehman, Gary J. Murray, Lisa A. Bowers, John A. Barranger

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49 Scopus citations

Abstract

Two cDNA clones (λGC-1 and λGC-2) for human β-glucocerebrosidase [EC 3.2.1.45] have been isolated from a human hepatoma library in λgtll by immunological screening using monospecific polyclonal antibody for β-glucocerebrosidase. Restriction endonuclease mapping indicates that these clones are probably identical in size, each with a 1900 bp insert. The 50 kDa size of the insert-encoded polypeptide produced by these clones in fusion with β-galactosidase of λgtll in E. coli BNN103 is consistent with the size of the nascent form of β-glucocerebrosidase. These fusion proteins are shown by Western blotting to react with antibody to β-glucocerebrosidase. Amino acid sequence deduced from the nucleotide sequence of the insert ir pGC-1 is identical to known amino acid sequence of β-glucocerebrosidase, and thus, confirms that the clones are specific for β-glucocerebrosidase.

Original languageEnglish (US)
Pages (from-to)574-580
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume123
Issue number2
DOIs
StatePublished - Sep 17 1984
Externally publishedYes

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ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Ginns, E. I., Choudary, P. V., Martin, B. M., Winfield, S., Stubblefield, B., Mayor, J., Merkle-Lehman, D., Murray, G. J., Bowers, L. A., & Barranger, J. A. (1984). Isolation of cDNA clones for human β-glucocerebrosidase using the λgtll expression system. Biochemical and Biophysical Research Communications, 123(2), 574-580. https://doi.org/10.1016/0006-291X(84)90268-7