The lac operon introduced into Proteus mirabilis shows two anomalies of expression: the maximal induced level is reduced by about an order of magnitude, and the basal level becomes about 100 times higher than in E. coli, as a result of which the induction ratio appears very small, around 2-5 in cont
Anomalous expression of the E. coli lac operon in Proteus mirabilis
β Scribed by Roberts, Margo ;Baumberg, Simon
- Publisher
- Springer
- Year
- 1984
- Tongue
- English
- Weight
- 772 KB
- Volume
- 198
- Category
- Article
- ISSN
- 0026-8925
No coin nor oath required. For personal study only.
β¦ Synopsis
The lac operon shows anomalous expression in Proteus mirabilis: the maximal induced level is 10% or less of that in E. coli, while repression reduces this by a factor of only 2-5. We have sought to determine whether this effect relates in any way to CRP-mediated activation of expression, by comparing expression in P. mirabilis of lac operons (introduced for technical reasons on IncP1 plasmids) either regulatorily wild-type or bearing L8 or L8UV5. Derivatives of RP1 bearing L8UV5 were obtained by homogenotisation of pGC9114 (RP1::Tn951) in a L8UV5 background; while derivatives of RP4 bearing lac+, L8 or L8UV5 were obtained by Mu-mediated translocation of chromosomal regions bearing these alleles, following partial heat-induction of Mucts62 on pGM14 (RP4::Mucts62) in the appropriate hosts. These plasmids could be readily transferred to, and stably maintained in, the P. mirabilis strains employed. It was found that L8 reduced the maximal level of beta-galactosidase activity, and L8UV5 restored this activity to around wild-type, in P. mirabilis quantitatively very much as in E. coli. Nevertheless, the low maximal level of expression and high basal level characteristic of the former host were unchanged. The simplest explanation of these results is that P. mirabilis contains a protein that mimics the E. coli CRP protein in interacting with the appropriate DNA binding site and thereby stimulating transcription; and that the anomalous regulation of lac in this host is unconnected with the CRP system.
π SIMILAR VOLUMES
We were kindly provided by Z. al-Doori (Dept. of Molecular Biology, University of Edinburgh) with a hybrid plasmid produced by in vitro techniques, pZD23 (A1- Doori et al. 1982). This plasmid is derived from pZD100, which consists of a 2drpoBC18 (=2drpoB3) prophage inserted into RP42att, by a deleti
The expression and stability of Escherichia coli F-primes in Proteus mirabilis is examined. It is possible to consecutively introduce, and stably maintain, the DNA of several E. coli F-primes in P. mirabilis in the absence of selective pressure for all or some of the plasmids. Additionally, we can r
Three temperature-sensitive mutant strains for RNA polymerase beta or beta' subunits (carrying mutations tsx, A2R7 and R120) were used in order to investigate the dependence of the induced lac expression on stimulation by cyclic AMP after the shift to non-permissive temperature. High temperature low
Various Escherichia coli K12 Hfr donors transfer at low frequency portions of the E. coli genome to Proteus mirabilis. By remating such Proteus hybrids with the same or a different E. coli Hfr strain, other genetic characters could be added to yield diploid Proteus hybrids which contained more than