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Construction and characterization of F plasmid‐based expression vectors
Author(s) -
Jones Kristala L.,
Keasling Jay D.
Publication year - 1998
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19980920)59:6<659::aid-bit1>3.0.co;2-e
Subject(s) - plasmid , computational biology , vector (molecular biology) , characterization (materials science) , expression vector , biology , chemistry , genetics , gene , recombinant dna , nanotechnology , materials science
A low‐copy expression vector has been constructed from a 9 Kbp region of the Escherichia coli F plasmid containing the oriV and oriS origins of replication. This plasmid carries the β‐lactamase gene (Ap r ) and the ara BAD promoter/ ara C regulator for arabinose‐inducible gene expression. A derivative which carries a lac Z reporter gene was found to be stably maintained for at least 150 generations. A related multi‐copy plasmid was stably maintained in arabinose‐free medium, but no plasmid‐bearing segregants remained after 60 generations when lac Z expression was induced. Induced expression resulted in 27% (multi‐copy) and 12% (low‐copy) decreases in growth rate. The uninduced levels of β‐galactosidase were 200 units (multi‐copy) and 15 units (low‐copy). © 1998 John‐Wiley & Sons, Inc. Biotechnol Bioeng 59:659–665, 1998.