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Expression of a pokeweed antiviral protein in Escherichia coli and its characterization
Author(s) -
Kataoka Jiro,
Ago Hideo,
Habuka Noriyuki,
Furuno Masahiro,
Masuta Chikara,
Miyano Masashi,
Koiwai Akira
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)81651-f
Subject(s) - escherichia coli , reticulocyte , ribosome , biochemistry , signal peptide , protein biosynthesis , biology , microbiology and biotechnology , rna , chemistry , recombinant dna , gene
Two expression vectors were constructed to produce a putative mature α‐pokeweed antiviral protein (α‐PAP) in Escherichia coli with its NH 2 ‐ and COOH‐terminal extrapeptides excised. One was for its intracellular expression with a methionine at its NH 2 ‐terminal. The other was for its secretion using an ompA signal peptide. The former product was purified from the total soluble proteins of the transformant with a yield of 1.74 mg/liter and the latter had a yield of 5.55 mg/liter. Both products exhibited RNA N ‐glycosidase activity on wheat ribosomes and inhibitory activity to protein synthesis in a rabbit reticulocyte system.