Stability and replication control of Escherichia coli minichromosomes
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Stability and replication control of Escherichia coli minichromosomes. / Løbner-Olesen, A; Atlung, T; Rasmussen, K V.
In: Journal of Bacteriology, Vol. 169, No. 6, 06.1987, p. 2835-42.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Stability and replication control of Escherichia coli minichromosomes
AU - Løbner-Olesen, A
AU - Atlung, T
AU - Rasmussen, K V
PY - 1987/6
Y1 - 1987/6
N2 - A stabilized minichromosome--a plasmid replicating from the chromosomal origin oriC--was constructed by cloning the sopA,B,C, genes from plasmid F. This minichromosome had a loss frequency of less than 10(-3), while that of the nonstabilized parental plasmid was 2 X 10(-2) to 4 X 10(-2). Both minichromosomes had the same average copy number per chromosomal origin, and the copy numbers were constant over an eightfold range of growth rates. Different mutations in the mioC gene and promoter, from which transcription enters oriC, were constructed, and their effects on minichromosome copy number and stability were tested. The results indicated that normal replication control at oriC was independent of the MioC protein and most of the sequences between the promoter and oriC, but required both transcription from the mioC promoter and probably also the presence of the DnaA box (DnaA protein-binding site) just upstream of the mioC promoter. Transcription from the mioC promoter was shown to be efficiently repressed in vivo after overproduction of DnaA protein and to be derepressed at the nonpermissive temperature in six different dnaA(Ts) mutants.
AB - A stabilized minichromosome--a plasmid replicating from the chromosomal origin oriC--was constructed by cloning the sopA,B,C, genes from plasmid F. This minichromosome had a loss frequency of less than 10(-3), while that of the nonstabilized parental plasmid was 2 X 10(-2) to 4 X 10(-2). Both minichromosomes had the same average copy number per chromosomal origin, and the copy numbers were constant over an eightfold range of growth rates. Different mutations in the mioC gene and promoter, from which transcription enters oriC, were constructed, and their effects on minichromosome copy number and stability were tested. The results indicated that normal replication control at oriC was independent of the MioC protein and most of the sequences between the promoter and oriC, but required both transcription from the mioC promoter and probably also the presence of the DnaA box (DnaA protein-binding site) just upstream of the mioC promoter. Transcription from the mioC promoter was shown to be efficiently repressed in vivo after overproduction of DnaA protein and to be derepressed at the nonpermissive temperature in six different dnaA(Ts) mutants.
KW - Bacterial Proteins/physiology
KW - Chromosomes, Bacterial/physiology
KW - DNA Replication
KW - Escherichia coli/genetics
KW - Gene Expression Regulation
KW - Genes, Bacterial
KW - Plasmids
KW - Promoter Regions, Genetic
KW - Transcription, Genetic
M3 - Journal article
C2 - 3294807
VL - 169
SP - 2835
EP - 2842
JO - Journal of Bacteriology
JF - Journal of Bacteriology
SN - 0021-9193
IS - 6
ER -
ID: 200973510