Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1990 Mar;172(3):1595–1599. doi: 10.1128/jb.172.3.1595-1599.1990

In vivo expression of the Pseudomonas stutzeri maltotetraose-forming amylase gene (amyP).

M Fujita 1, M Futai 1, A Amemura 1
PMCID: PMC208637  PMID: 1689717

Abstract

Northern hybridization and S1 nuclease mapping revealed that the amyP gene coding for maltotetraose-forming amylase of Pseudomonas stutzeri MO-19 is transcribed as a monocistronic mRNA of 2.0 kilobases and that the transcription start site is located 81 base pairs upstream from the first nucleotide of the initiation codon. The amyP gene was expressed weakly in Escherichia coli, and transcription started 49 base pairs downstream of the P. stutzeri MO-19 transcription start site. Synthesis of the amylase in P. stutzeri MO-19 was induced by the addition of maltose to the culture medium and was repressed by the addition of glucose. The induction by maltose was shown to be result of transcription induction of the amyP gene. In contrast, glucose did not repress transcription initiation of amyP, indicating that it controls synthesis of the enzyme, probably at the posttranscriptional level.

Full text

PDF
1595

Images in this article

1596FIG. 2
on p.1596
1597FIG. 3
on p.1597
1599FIG. 7
on p.1599

Selected References

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

  1. Aiba H., Adhya S., de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [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. Friedman D. I., Olson E. R., Georgopoulos C., Tilly K., Herskowitz I., Banuett F. Interactions of bacteriophage and host macromolecules in the growth of bacteriophage lambda. Microbiol Rev. 1984 Dec;48(4):299–325. doi: 10.1128/mr.48.4.299-325.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fujita M., Amemura A., Futai M. Transcription of the isoamylase gene (iam) in Pseudomonas amyloderamosa SB-15. J Bacteriol. 1989 Aug;171(8):4320–4325. doi: 10.1128/jb.171.8.4320-4325.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fujita M., Torigoe K., Nakada T., Tsusaki K., Kubota M., Sakai S., Tsujisaka Y. Cloning and nucleotide sequence of the gene (amyP) for maltotetraose-forming amylase from Pseudomonas stutzeri MO-19. J Bacteriol. 1989 Mar;171(3):1333–1339. doi: 10.1128/jb.171.3.1333-1339.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hirschman J., Wong P. K., Sei K., Keener J., Kustu S. Products of nitrogen regulatory genes ntrA and ntrC of enteric bacteria activate glnA transcription in vitro: evidence that the ntrA product is a sigma factor. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7525–7529. doi: 10.1073/pnas.82.22.7525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hunt T. P., Magasanik B. Transcription of glnA by purified Escherichia coli components: core RNA polymerase and the products of glnF, glnG, and glnL. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8453–8457. doi: 10.1073/pnas.82.24.8453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Inouye S., Nakazawa A., Nakazawa T. Expression of the regulatory gene xylS on the TOL plasmid is positively controlled by the xylR gene product. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5182–5186. doi: 10.1073/pnas.84.15.5182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Inouye S., Nakazawa A., Nakazawa T. Molecular cloning of regulatory gene xylR and operator-promoter regions of the xylABC and xylDEGF operons of the TOL plasmid. J Bacteriol. 1983 Sep;155(3):1192–1199. doi: 10.1128/jb.155.3.1192-1199.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nicholson W. L., Chambliss G. H. Isolation and characterization of a cis-acting mutation conferring catabolite repression resistance to alpha-amylase synthesis in Bacillus subtilis. J Bacteriol. 1985 Mar;161(3):875–881. doi: 10.1128/jb.161.3.875-881.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Olson E. R., Flamm E. L., Friedman D. I. Analysis of nutR: a region of phage lambda required for antitermination of transcription. Cell. 1982 Nov;31(1):61–70. doi: 10.1016/0092-8674(82)90405-6. [DOI] [PubMed] [Google Scholar]
  12. Raibaud O., Richet E. Maltotriose is the inducer of the maltose regulon of Escherichia coli. J Bacteriol. 1987 Jul;169(7):3059–3061. doi: 10.1128/jb.169.7.3059-3061.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Robyt J. F., Ackerman R. J. Isolation, purification, and characterization of a maltotetraose-producing amylase from Pseudomonas stutzeri. Arch Biochem Biophys. 1971 Jul;145(1):105–114. doi: 10.1016/0003-9861(71)90015-4. [DOI] [PubMed] [Google Scholar]
  14. Rothstein D. M., Devlin P. E., Cate R. L. Expression of alpha-amylase in Bacillus licheniformis. J Bacteriol. 1986 Nov;168(2):839–842. doi: 10.1128/jb.168.2.839-842.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Shine J., Dalgarno L. Determinant of cistron specificity in bacterial ribosomes. Nature. 1975 Mar 6;254(5495):34–38. doi: 10.1038/254034a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES