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Expression of the Escherichia coli ftsZ gene: trials and tribulations of gene fusion studies

Published online by Cambridge University Press:  14 April 2009

Aline Robin  and
Richard D'Ari
Aline Robin*
Affiliation:
Institut Jacques Monod, C.N.R.S., Université Paris 7, 2, place Jussieu, 75251 Paris Cedex 05, France
Richard D'Ari
Affiliation:
Institut Jacques Monod, C.N.R.S., Université Paris 7, 2, place Jussieu, 75251 Paris Cedex 05, France
*
* Corresponding author.
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The ftsZ gene of Escherichia coli, which codes for an essential cell division protein, is subjected to multiple regulation, as shown in part with studies using an ftsZ::lacZ operon fusion located on phage λJFLIOO. Using this same fusion, we sought to isolate regulatory mutants overexpressing ftsZ by selecting mutants able to grow on lactose. One Lac+ mutant was obtained which overexpressed the ftsZ::lacZ fusion 70-fold. The mutation responsible for the overexpression lies in a new gene, cot, located near 56 min on the E. coli genetic map. The cot mutation probably affects the transcription of a chromosomal open reading frame, 0RF1, lying downstream of the bioA gene and adjacent to the ftzZ::lacZ fusion of the λJFL100 prophage integrated at attλ. Using an ftsZ84(Ts) strain, in which there was a double selection for overexpression of both ftsZ::lacZ and ftsZ+, no Lac+Tr mutants were obtained from 3·6 × 1010 bacteria; the introduction of a mutL allele, increasing spontaneous base substitution mutation rates 75-fold, did not permit us to isolate such a mutant. We conclude that Lac+ftsZ-constitutive mutations cannot be obtained in λJFL100 lysogens by a single base substitution.

Type
Research Article
Information
Genetics Research , Volume 61 , Issue 1 , February 1993 , pp. 1 - 8
Copyright
Copyright © Cambridge University Press 1993

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