Effects of the par locus on the growth rate and structural stability of recombinant cells.
A decreased growth rate of recombinant cells is observed when a cloned gene protein encoded in a multicopy plasmid is induced from a strong promoter. This negative effect on the cell growth rate may be the consequence of alternate use or reallocation of energy, precursor metabolites, and protein synthesizing machinery. In order to analyze the causes for this adverse effect, we have studied how genetic elements present in multicopy plasmids affect the growth of recombinant Escherichia coli (K12 delta H1 delta trpEA). Turning on of the PL promoter, whether or not protein-coding sequences were present downstream of the promoter, decreased the growth of the recombinant cells by 15-50%. This result suggests that the essential factor(s) for cell growth may be sequestered by the genetic elements present in the plasmids and thus may cause the decreased growth. The negative effect of the de-repressed PL promoters on cell growth was partially reversed when the par sequence from pSC101 was inserted into the same plasmid. In each case when the plasmid carried only the PL promoter, the PL promoter followed by the N-terminal part of the protein-coding region, or the PL promoter followed by complete sequences encoding a protein, the par sequence exhibited a positive effect on the growth rate of the recombinant cells, which usually grow at a slower rate than the host cell.(ABSTRACT TRUNCATED AT 250 WORDS)
Kim
J Y
JY
Microbiology Graduate Group, School of Medicine, University of California at Davis 95616.
Kang
H A
HA
Ryu
D D
DD
D001483:Base Sequence; D001665:Binding Sites; D002455:Cell Division / Q000502:physiology; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development*; D005798:Genes, Bacterial / Q000235:genetics / Q000502:physiology; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics* / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology; D011994:Recombinant Proteins / Q000235:genetics; D012270:Ribosomes / Q000378:metabolism
D004274:DNA, Recombinant; D011994:Recombinant Proteins