Open AccessSynthesis and overproduction of the 5A protein of insertion sequence IS5
Author(s) -
Jeffrey M. Chernak,
E J Schlaffer,
Hamilton O. Smith
Publication year - 1988
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.170.11.5368-5370.1988
Subject(s) - biology , overproduction , plasmid , escherichia coli , microbiology and biotechnology , dna , in vitro , open reading frame , peptide sequence , genetics , gene
We have demonstrated both the synthesis and overproduction of the 5A protein encoded by the longest open reading frame of the bacterial insertion sequence IS5. Expression was obtained in vitro and in Escherichia coli maxicells from plasmids containing IS5 in either orientation, as well as in vitro from a restriction fragment containing exclusively IS5 DNA. When IS5 was cloned in the appropriate orientation downstream of a strong tac promoter, production of the 5A protein was increased to 10 to 20% of the total protein synthesized in vitro.