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

doi:10.1016/0006-291X(84)90268-7

Isolation of cDNA clones for human β-glucocerebrosidase using the λ_gtll 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

Research output: Contribution to journal › Article

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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 λ_gtll 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 λ_gtll 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)
Pages (from-to)	574-580
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	123
Issue number	2
DOIs	https://doi.org/10.1016/0006-291X(84)90268-7
State	Published - Sep 17 1984
Externally published	Yes

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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 λ_gtll expression system. Biochemical and Biophysical Research Communications, 123(2), 574-580. https://doi.org/10.1016/0006-291X(84)90268-7

Isolation of cDNA clones for human β-glucocerebrosidase using the λ_gtll expression system. / Ginns, Edward I.; Choudary, Prabhakara V; Martin, Brian M.; Winfield, Suzanne; Stubblefield, Barbara; Mayor, June; Merkle-Lehman, Denise; Murray, Gary J.; Bowers, Lisa A.; Barranger, John A.

In: Biochemical and Biophysical Research Communications, Vol. 123, No. 2, 17.09.1984, p. 574-580.

Research output: Contribution to journal › Article

Ginns, Edward I. ; Choudary, Prabhakara V ; Martin, Brian M. ; Winfield, Suzanne ; Stubblefield, Barbara ; Mayor, June ; Merkle-Lehman, Denise ; Murray, Gary J. ; Bowers, Lisa A. ; Barranger, John A. / Isolation of cDNA clones for human β-glucocerebrosidase using the λ_gtll expression system. In: Biochemical and Biophysical Research Communications. 1984 ; Vol. 123, No. 2. pp. 574-580.

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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.",

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10.1016/0006-291X(84)90268-7