Advertisement

Applied Microbiology and Biotechnology

, Volume 75, Issue 5, pp 1127–1132 | Cite as

Identification of promoters of two dehydrogenase genes in Ketogulonicigenium vulgare DSM 4025 and their strength comparison in K. vulgare and Escherichia coli

  • Shulin Fu
  • Weicai ZhangEmail author
  • Aiguang Guo
  • Jianhua Wang
Applied Genetics and Molecular Biotechnology

Abstract

Promoters of sorbose dehydrogenase gene sdh and sorbosone dehydrogenase gene sndh (Psdh and Psndh) in Ketogulonicigenium vulgare DSM 4025 were identified. The transcription initiation site (TIS) of Psdh was guanine 74 bp upstream of the start codon of sdh and the TIS of Psndh was adenine 113 bp upstream of the first codon of sndh. Comparing Psdh and Psndh, consensus sequences were found, which were TAVCVT (V=A, C or G) and THGAHC (H=A, C or T) for their putative −10 and −35 regions, respectively, and the spans between the 2 regions were 17 bp. Psdh and Psndh promoters may be constitutive in K. vulgare DSM 4025 when cultured in HJ medium. Semiquantitative RT-PCR analysis showed that the Psdh promoter was about 2.5 times stronger than Psndh in strength in K. vulgare DSM 4025. In Escherichia coli, Psdh and Psndh demonstrated strong activity with the former about two times stronger than the latter. DCIP decoloration method and reporter plasmids pSDH or pSNDH may be applied to discover promoters of genes in E. coli and to determine their strength in one step.

Keywords

Promoters Determination Strength comparison Ketogulonicigenium vulgare DSM 4025 E. coli 

Notes

Acknowledgement

We are grateful to Professor Susheng Yang for providing vector pBBR1MCS2. This work is supported by the National Natural Science Foundation of China (No. 30470058).

References

  1. Davis BJ (1964) Disc electrophoresis. II. Method and application to human serum proteins. Ann N Y Acad Sci 121:404–427PubMedCrossRefGoogle Scholar
  2. Delany I, Rappuoli R, Scarlato V (2004) Fur functions as an activator and as a repressor of putative virulence genes in Neisseria meningitidis. Mol Microbiol 52:1081–1090PubMedCrossRefGoogle Scholar
  3. Fujishige K, Kotera J, Yuasa K, Omori K (2000) The human phosphodiesterase PDE10A gene genomic organization and evolutionary relatedness with other PDEs containing GAF domains. Eur J Biochem 267:5943–5951PubMedCrossRefGoogle Scholar
  4. Gupta A, Phung LT, Taylor DE, Silver S (2001) Diversity of silver resistance genes in IncH incompatibility group plasmids. Microbiology 147:3393–3402PubMedGoogle Scholar
  5. Jacob D, Lewin A, Meister B, Appel B (2002) Plant-specific promoter sequences carry elements that are recognized by the eubacterial transcription machinery. Transgenic Res 11:291–303PubMedCrossRefGoogle Scholar
  6. Knauf VC, Nester EW (1982) Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid 8:45–54PubMedCrossRefGoogle Scholar
  7. Leblanc C, Falciatore A, Watanabe M, Bowler C (1999) Semi-quantitative RT-PCR analysis of photoregulated gene expression in marine diatoms. Plant Mol Biol 40:1031–1044PubMedCrossRefGoogle Scholar
  8. Lewin A, Mayer M, Chusainow J, Jacob D, Appel B (2005) Viral promoters can initiate expression of toxin genes introduced into Escherichia coli. BMC Biotechnol 5:19PubMedCrossRefGoogle Scholar
  9. Master S, Zahrt TC, Song J, Deretic V (2001) Mapping of Mycobacterium tuberculosis katG promoters and their differential expression in infected macrophages. J Bacteriol 183:4033–4039PubMedCrossRefGoogle Scholar
  10. Michael E, Kovach, Philip H, Elzer (1995) Four new derivatives of the broad-host-range cloning vector pBBR1mcs, carrying different antibiotic-resistance cassettes. Gene 166:175–176CrossRefGoogle Scholar
  11. Miyazaki T, Sugisawa T, Hoshino T (2006) Pyrroloquinoline quinone-dependent dehydrogenases from Ketogulonicigenium vulgare catalyze the direct conversion of l-sorbosone to l-ascorbic acid. Appl Environ Microbiol 72:1487–1495PubMedCrossRefGoogle Scholar
  12. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
  13. Shinjoh M, Tomiyama N, Asakura A, Hoshino T (1995) Cloning and nucleotide sequencing of the membrane-bound l-sorbosone dehydrogenase gene of Acetobacter liquefaciens IFO 12258 and its expression in Gluconobacter oxydans. Appl Environ Microbiol 61:413–420PubMedGoogle Scholar
  14. Sugisawa T, Miyazaki T, Hoshino T (2005) Microbial production of l-ascorbic acid from d-sorbitol, l-sorbose, l-glucose and l-sorbosone by Ketogulonicigenium vulgare DSM 4025. Biosci Biotechnol Biochem 69:659–662PubMedCrossRefGoogle Scholar
  15. Takahashi H, Fujiwara H (1999) Transcription analysis of the telomeric repeat-specific retrotransposons TRAS1 and SART1 of the silkworm Bombyx mori. Nucleic Acids Res 27:2015–2021PubMedCrossRefGoogle Scholar
  16. Urbance JW, Bratina BJ, Stoddard SF, Schmidt TM (2001) Taxonomic characterization of Ketogulonigenium vulgare gen. nov., sp. nov. and Ketogulonigenium robustum sp. nov., which oxidize l-sorbose to 2-keto-l-gulonic acid. Int J Syst Evol Microbiol 51:1059–1070PubMedGoogle Scholar
  17. Xiao HS, Huang QH, Zhang FX, Bao L, Lu YJ et al (2002) Identification of gene expression profile of dorsal root ganglion in the rat peripheral axotomy model of neuropathic pain. Proc Natl Acad Sci USA 99:8360–8365PubMedCrossRefADSGoogle Scholar
  18. Zhang WC, Jiao YH, Yuan HJ, Xie L (2003) A novel l-sorbose dehydrogenase gene and the encoding protein. China Patent, CN03102060.7Google Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Shulin Fu
    • 1
    • 2
  • Weicai Zhang
    • 2
    Email author
  • Aiguang Guo
    • 1
  • Jianhua Wang
    • 2
  1. 1.College of Life ScienceNorthwest Sci-Tec University of Agriculture and ForestryYanglingPeople’s Republic of China
  2. 2.Beijing Institute of BiotechnologyBeijingPeople’s Republic of China

Personalised recommendations