A phycocyanin-deficient mutant of synechocystis PCC 6714 with a single-base substitution upstream of the cpc operon.
The structure and expression of the cpc operon encoding phycocyanin subunits and linker polypeptides in a phycocyanin-deficient mutant (PD-1) and the wild-type of Synechocystis PCC 6714 were analyzed. The results of sequence and Northern blot analyses of the wild type indicate that the cpc operon consists of cpcB, cpcA, cpcC1, cpcC2 and cpcD, in that order. The levels of the transcripts in PD-1 were one-tenth to one-sixth as high as those in the wild type. In the PD-1 genome, a single-base substitution of C for T has occurred at base 259 upstream of the translational initiation codon of cpcB (at three bases downstream of the putative -10 region). To evaluate the in vivo transcription activities of these promoters in a cyanobacterium, we constructed vectors for the transformation of Synechococcus PCC7942, pANY1 and pANY2, which contain the upstream region of cpcB of the wild type (pANY1) or PD-1 (pANY2) and the promoter-less luxAB fusion. The bioluminescence of the transformants with pANY2 was one-tenth to one-sixth as high as that with pANY1. The coincidence of the results of Northern analysis and the promoter assay shows that the phycocyanin deficiency of PD-1 is due to the single-base substitution in the upstream region of the cpc operon.
Nakajima
Y
Y
Advanced Technology Research Center, Mitsubishi Heavy Industries, Ltd., Yokohama, 236-8515 Japan. nakajima@atrc.mhi.co.jp
Fujiwara
S
S
Sawai
H
H
Imashimizu
M
M
Tsuzuki
M
M
D001483:Base Sequence; D002460:Cell Line; D000458:Cyanobacteria / Q000235:genetics*; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D009154:Mutation; D009876:Operon / Q000235:genetics; D010798:Phycocyanin / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D016415:Sequence Alignment
D010798:Phycocyanin