Construction of lacZ promoter probe vectors for use in Synechococcus: application to the identification of CO2-regulated promoters.
It was shown that the Escherichia coli lacZ gene could be expressed in the cyanobacterium Synechococcus R2 PCC7942 both as a plasmid-borne form and also integrated into the chromosome. A promoterless form of the lacZ gene was constructed and used as a reporter gene to make transcriptional fusions with cyanobacterial promoters using a shuttle vector system and also via a process of integration by homologous recombination. Synechococcus R2 promoter-lacZ gene fusions were then used to identify CO2-regulated promoters, by quantitatively assessing beta-galactosidase activity under high and low CO2 conditions using a fluorescence assay. Several promoters induced under low CO2 conditions were detected.
Scanlan
D J
DJ
Department of Biological Sciences, University of Warwick, Coventry, U.K.
Bloye
S A
SA
Mann
N H
NH
Hodgson
D A
DA
Carr
N G
NG
D000595:Amino Acid Sequence; D001483:Base Sequence; D002245:Carbon Dioxide; D003001:Cloning, Molecular / Q000379:methods; D000458:Cyanobacteria / Q000235:genetics*; D005822:Genetic Vectors; D007763:Lac Operon; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic; D001616:beta-Galactosidase / Q000235:genetics
D002245:Carbon Dioxide; D001616:beta-Galactosidase