Abstract
An expression vector, pFC1, has been constructed based on the promiscuous plasmid RSF1010, which provides autonomous replication in several cyanobacteria of the generaSynechocystis andSynechococcus [Mermet-Bouvier et al., Curr Microbiol 26:323–327]. pFC1 harbors the λcI857 repressor-encoding gene andp R promoter, followed by the λcro ribosome-binding site and ATG translation initiation codon. The latter is located within the uniqueNdeI restriction site (CATATG) of pFC1 and can be exposed after cleavage with this enzyme for in-frame fusion with the protein-coding sequence to be expressed. TheEscherichia coli lacZ reporter gene cloned in pFC1 appeared to be highly expressed in heat-inducedE. coli or cyanobacterial cells. In every case, β-galactosidase amounted to at least 10% of soluble proteins.
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Mermet-Bouvier, P., Chauvat, F. A conditional expression vector for the cyanobacteriaSynechocystis sp. strains PCC6803 and PCC6714 orSynechococcus sp. strains PCC7942 and PCC6301. Current Microbiology 28, 145–148 (1994). https://doi.org/10.1007/BF01571055
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DOI: https://doi.org/10.1007/BF01571055