Expression of theEscherichia coli ftsZgene: trials and tribulations of gene fusion studies
SummaryTheftsZgene ofEscherichia coli, which codes for an essential cell division protein, is subjected to multiple regulation, as shown in part with studies using anftsZ::lacZoperon fusion located on phage λJFLIOO. Using this same fusion, we sought to isolate regulatory mutants overexpressingftsZby selecting mutants able to grow on lactose. One Lac+mutant was obtained which overexpressed theftsZ::lacZfusion 70-fold. The mutation responsible for the overexpression lies in a new gene,cot, located near 56 min on theE. coligenetic map. Thecotmutation probably affects the transcription of a chromosomal open reading frame, 0RF1, lying downstream of thebioAgene and adjacent to theftzZ::lacZfusion of the λJFL100 prophage integrated atattλ. Using anftsZ84(Ts) strain, in which there was a double selection for overexpression of bothftsZ::lacZandftsZ+, no Lac+Tr mutants were obtained from 3·6 × 1010bacteria; the introduction of amutLallele, 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.