A constitutive expression vector system driven by the deo P1P2 promoters of Escherichia coli.
The P1P2 promoters of Escherichia coli K12 deo operon, residing on an AvaII restriction fragment, were used to construct a new expression vector. To evaluate the potential of the P1P2-driven expression system we have inserted the sequence of human superoxide dismutase (hSOD) downstream of the deo ribosome binding site. Expression of hSOD was evaluated by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis and enzyme activity. In crude cell extracts hSOD expression levels were found to be high in hosts possessing no deoR or cytR repressors. Highest levels of hSOD expression were obtained with a high-copy-number plasmid regardless of the host used. Expressed hSOD can account for 35%-40% of total protein in E. coli.
Department of Molecular Biology, Biotechnology General Inc., Rehovot, Israel.
M
M
Fischer
S
S
Fytlovitch
B
B
Amit
A
A
Wortzel
Y
Y
Beck
D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D009876:Operon; D010957:Plasmids; D011401:Promoter Regions, Genetic; D013379:Substrate Specificity; D013482:Superoxide Dismutase / Q000235:genetics*
D013482:Superoxide Dismutase