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. 1981 Nov;148(2):413–418. doi: 10.1128/jb.148.2.413-418.1981

Molecular cloning of gene xylS of the TOL plasmid: evidence for positive regulation of the xylDEGF operon by xylS.

S Inouye, A Nakazawa, T Nakazawa
PMCID: PMC216221  PMID: 6271729

Abstract

The xylDEGF operon and the regulatory gene xylS of the TOL plasmid found in Pseudomonas putida mt-2 were cloned onto Escherichia coli vector plasmids. A 9.5-kilobase fragment, derived from the TOL segment of pTN2 deoxyribonucleic acid, carried the xyl genes D, E, G, and F, which encode toluate oxygenase, catechol 2,3-oxygenase, 2-hydroxymuconic semialdehyde dehydrogenase, and 2-hydroxymuconic semialdehyde hydrolase, respectively. The enzymes were noninducible unless a 3-kilobase PstI fragment, derived also from the TOL segment, was provided in either cis or trans. The PstI fragment appeared to contain the regulatory gene xylS, which produced a positive regulator. The regulator was activated by m-toluate or benzoate, but not by m-xylene or m-methylbenzyl alcohol. the map positions of xylG and xylF were also determined.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  4. Chakrabarty A. M., Friello D. A., Bopp L. H. Transposition of plasmid DNA segments specifying hydrocarbon degradation and their expression in various microorganisms. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3109–3112. doi: 10.1073/pnas.75.7.3109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Downing R., Broda P. A cleavage map of the TOL plasmid of Pseudomonas putida mt-2. Mol Gen Genet. 1979;177(1):189–191. doi: 10.1007/BF00267270. [DOI] [PubMed] [Google Scholar]
  7. Débarbouillé M., Shuman H. A., Silhavy T. J., Schwartz M. Dominant constitutive mutations in malT, the positive regulator gene of the maltose regulon in Escherichia coli. J Mol Biol. 1978 Sep 15;124(2):359–371. doi: 10.1016/0022-2836(78)90304-2. [DOI] [PubMed] [Google Scholar]
  8. Englesberg E., Wilcox G. Regulation: positive control. Annu Rev Genet. 1974;8:219–242. doi: 10.1146/annurev.ge.08.120174.001251. [DOI] [PubMed] [Google Scholar]
  9. Franklin F. C., Williams P. A. Construction of a partial diploid for the degradative pathway encoded by the TOL plasmid (pWWO) from Pseudomonas putida mt-2: evidence for the positive nature of the regulation by the xyIR gene. Mol Gen Genet. 1980 Jan;177(2):321–328. doi: 10.1007/BF00267445. [DOI] [PubMed] [Google Scholar]
  10. Hedges R. W., Jacob A. E., Crawford I. P. Wide ranging plasmid bearing the Pseudomonas aeruginosa tryptophan synthase genes. Nature. 1977 May 19;267(5608):283–284. doi: 10.1038/267283a0. [DOI] [PubMed] [Google Scholar]
  11. Inouye S., Nakazawa A., Nakazawa T. Molecular cloning of TOL genes xylB and xylE in Escherichia coli. J Bacteriol. 1981 Mar;145(3):1137–1143. doi: 10.1128/jb.145.3.1137-1143.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Nakazawa T., Hayashi E., Yokota T., Ebina Y., Nakazawa A. Isolation of TOL and RP4 recombinants by integrative suppression. J Bacteriol. 1978 Apr;134(1):270–277. doi: 10.1128/jb.134.1.270-277.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nakazawa T., Inouye S., Nakazawa A. Physical and functional mapping of RP4-TOL plasmid recombinants: analysis of insertion and deletion mutants. J Bacteriol. 1980 Oct;144(1):222–231. doi: 10.1128/jb.144.1.222-231.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nakazawa T., Yokota T. Benzoate metabolism in Pseudomonas putida(arvilla) mt-2: demonstration of two benzoate pathways. J Bacteriol. 1973 Jul;115(1):262–267. doi: 10.1128/jb.115.1.262-267.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ogden S., Haggerty D., Stoner C. M., Kolodrubetz D., Schleif R. The Escherichia coli L-arabinose operon: binding sites of the regulatory proteins and a mechanism of positive and negative regulation. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3346–3350. doi: 10.1073/pnas.77.6.3346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Worsey M. J., Franklin F. C., Williams P. A. Regulation of the degradative pathway enzymes coded for by the TOL plasmid (pWWO) from Pseudomonas putida mt-2. J Bacteriol. 1978 Jun;134(3):757–764. doi: 10.1128/jb.134.3.757-764.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Worsey M. J., Williams P. A. Metabolism of toluene and xylenes by Pseudomonas (putida (arvilla) mt-2: evidence for a new function of the TOL plasmid. J Bacteriol. 1975 Oct;124(1):7–13. doi: 10.1128/jb.124.1.7-13.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

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