A promoter-probe vector-host system for the cyanobacterium, Synechocystis PCC6803.
A vector-host system for testing promoters in the cyanobacterium Synechocystis PCC6803 has been constructed. It relies on a small Escherichia coli promoter-probe plasmid, pFF11, which has four unique restriction sites in a polylinker upstream from the cat reporter gene. This plasmid is able to obtain a cyanobacterial origin of replication by homologous recombination with the resident plasmid of the recipient host, generating a new E. coli-Synechocystis PCC6803 shuttle vector. This plasmid does not confer any detectable chloramphenicol acetyl transferase activity to this cyanobacterium in the absence of a promoter insert. Several heterologous promoters were tested in Synechocystis PCC6803 using this system. Results obtained with the lambda pR promoter and the repressor-encoding cI857 gene demonstrate that these elements can be used for high-level and tightly regulated gene expression in Synechocystis PCC6803.
Ferino
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F
Département de Biologie, C.E.N. Saclay, Gif-sur-Yvette, France.
Chauvat
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F
D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014170:Transformation, Genetic
D015500:Chloramphenicol O-Acetyltransferase