Amplification of ColE1 related plasmids in recombinant cultures of Escherichia coli after IPTG induction.
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Amplification of ColE1 related plasmids in recombinant cultures of Escherichia coli after IPTG induction. / Teich, A; Lin, Hongying; Andersson, L; Meyer, S; Neubauer, P.
in: J BIOTECHNOL, Jahrgang 64, Nr. 2-3, 2-3, 1998, S. 197-210.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Amplification of ColE1 related plasmids in recombinant cultures of Escherichia coli after IPTG induction.
AU - Teich, A
AU - Lin, Hongying
AU - Andersson, L
AU - Meyer, S
AU - Neubauer, P
PY - 1998
Y1 - 1998
N2 - ColE1-derived plasmids containing different recombinant genes which are controlled by the tac promoter were amplified following induction with IPTG, but no amplification occurred if product formation was not induced. The plasmid copy number of recombinant E. coli increased three- to sixfold within a period of about 6 h in shake flask experiments, batch cultures, and glucose-limited fed-batch cultivations. Plasmid amplification occurred in E. coli B strains as well as in K-12 strains with different plasmids (rop+ and rop-) coding for various heterologous proteins. The amplification was not caused by a toxic effect of IPTG, but was related to a strong inhibition of translation and chromosomal replication after the induction of heterologous gene expression. Similar to the amplification after chloramphenicol addition, plasmid replication proceeded even if oriC replication and translation were inhibited following strong induction of a recombinant gene. In accordance with the effect of chloramphenicol, the level of ppGpp, which is a negative regulator of ColE1 derived plasmid replication, decreased after induction.
AB - ColE1-derived plasmids containing different recombinant genes which are controlled by the tac promoter were amplified following induction with IPTG, but no amplification occurred if product formation was not induced. The plasmid copy number of recombinant E. coli increased three- to sixfold within a period of about 6 h in shake flask experiments, batch cultures, and glucose-limited fed-batch cultivations. Plasmid amplification occurred in E. coli B strains as well as in K-12 strains with different plasmids (rop+ and rop-) coding for various heterologous proteins. The amplification was not caused by a toxic effect of IPTG, but was related to a strong inhibition of translation and chromosomal replication after the induction of heterologous gene expression. Similar to the amplification after chloramphenicol addition, plasmid replication proceeded even if oriC replication and translation were inhibited following strong induction of a recombinant gene. In accordance with the effect of chloramphenicol, the level of ppGpp, which is a negative regulator of ColE1 derived plasmid replication, decreased after induction.
M3 - SCORING: Zeitschriftenaufsatz
VL - 64
SP - 197
EP - 210
JO - J BIOTECHNOL
JF - J BIOTECHNOL
SN - 0168-1656
IS - 2-3
M1 - 2-3
ER -
