Two cDNA clones (λGC-1 and λGC-2) for human β-glucocerebrosidase [EC 220.127.116.11] 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 language||English (US)|
|Number of pages||7|
|Journal||Biochemical and Biophysical Research Communications|
|State||Published - Sep 17 1984|
ASJC Scopus subject areas
- Molecular Biology