TY - JOUR
T1 - Isolation of cDNA clones for human β-glucocerebrosidase using the λgtll expression system
AU - Ginns, Edward I.
AU - Choudary, Prabhakara V
AU - Martin, Brian M.
AU - Winfield, Suzanne
AU - Stubblefield, Barbara
AU - Mayor, June
AU - Merkle-Lehman, Denise
AU - Murray, Gary J.
AU - Bowers, Lisa A.
AU - Barranger, John A.
PY - 1984/9/17
Y1 - 1984/9/17
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0021123304&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0021123304&partnerID=8YFLogxK
U2 - 10.1016/0006-291X(84)90268-7
DO - 10.1016/0006-291X(84)90268-7
M3 - Article
C2 - 6091633
AN - SCOPUS:0021123304
VL - 123
SP - 574
EP - 580
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 2
ER -