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. 1992 Jan;174(2):426–433. doi: 10.1128/jb.174.2.426-433.1992

mRNA analysis of the adc gene region of Clostridium acetobutylicum during the shift to solventogenesis.

U Gerischer 1, P Dürre 1
PMCID: PMC205733  PMID: 1370288

Abstract

By using primer extension analysis, we located the transcription start point of the acetoacetate decarboxylase (adc) gene of Clostridium acetobutylicum 90 nucleotides upstream from the initiation codon with A as the first transcribed nucleotide. From this site the promoter structure TTTACT(18 bp)TATAAT was identified; it shows high homology to the consensus sequences of gram-positive bacteria and Escherichia coli. Northern blot experiments revealed a length of 850 bases for the transcript of the adc gene. It thus represents a monocistronic operon. Transcription of adc was induced by conditions necessary for the onset of solvent formation. Induction occurred long before the respective fermentation product (acetone) could be detected in the medium. Transcription of the operon containing the genes for acetoacetyl coenzyme A:acetate/butyrate:coenzyme A transferase (designated ctf) downstream of the adc gene but divergently transcribed is also induced by conditions necessary for the onset of solvent formation. The length of the respective RNA transcript, 4.1 kb, indicates additional coding capacity, since the genes for the two subunits of the coenzyme A transferase cover only approximately 1.5 kb. No distinct transcripts for the other open reading frames of the adc gene region, ORF1 and ORF2, could be detected. Computer analysis indicated that ORF1, which showed significant similarity to the alpha-amylase gene of Bacillus subtilis (U. Gerischer and P. Dürre, J. Bacteriol. 172:6907-6918, 1990), probably is indeed a coding region. ORF2, however, does not seem to have a coding function.

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Selected References

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

  1. Cary J. W., Petersen D. J., Papoutsakis E. T., Bennett G. N. Cloning and expression of Clostridium acetobutylicum ATCC 824 acetoacetyl-coenzyme A:acetate/butyrate:coenzyme A-transferase in Escherichia coli. Appl Environ Microbiol. 1990 Jun;56(6):1576–1583. doi: 10.1128/aem.56.6.1576-1583.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Garnier T., Saurin W., Cole S. T. Molecular characterization of the resolvase gene, res, carried by a multicopy plasmid from Clostridium perfringens: common evolutionary origin for prokaryotic site-specific recombinases. Mol Microbiol. 1987 Nov;1(3):371–376. doi: 10.1111/j.1365-2958.1987.tb01944.x. [DOI] [PubMed] [Google Scholar]
  4. Gerischer U., Dürre P. Cloning, sequencing, and molecular analysis of the acetoacetate decarboxylase gene region from Clostridium acetobutylicum. J Bacteriol. 1990 Dec;172(12):6907–6918. doi: 10.1128/jb.172.12.6907-6918.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Graves M. C., Rabinowitz J. C. In vivo and in vitro transcription of the Clostridium pasteurianum ferredoxin gene. Evidence for "extended" promoter elements in gram-positive organisms. J Biol Chem. 1986 Aug 25;261(24):11409–11415. [PubMed] [Google Scholar]
  6. Helmann J. D., Chamberlin M. J. Structure and function of bacterial sigma factors. Annu Rev Biochem. 1988;57:839–872. doi: 10.1146/annurev.bi.57.070188.004203. [DOI] [PubMed] [Google Scholar]
  7. Janssen P. J., Jones D. T., Woods D. R. Studies on Clostridium acetobutylicum glnA promoters and antisense RNA. Mol Microbiol. 1990 Sep;4(9):1575–1583. [PubMed] [Google Scholar]
  8. Janssen P. J., Jones W. A., Jones D. T., Woods D. R. Molecular analysis and regulation of the glnA gene of the gram-positive anaerobe Clostridium acetobutylicum. J Bacteriol. 1988 Jan;170(1):400–408. doi: 10.1128/jb.170.1.400-408.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jones D. T., Woods D. R. Acetone-butanol fermentation revisited. Microbiol Rev. 1986 Dec;50(4):484–524. doi: 10.1128/mr.50.4.484-524.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kellmann J. W., Pichersky E., Piechulla B. Analysis of the diurnal expression patterns of the tomato chlorophyll a/b binding protein genes. Influence of light and characterization of the gene family. Photochem Photobiol. 1990 Jul;52(1):35–41. doi: 10.1111/j.1751-1097.1990.tb01752.x. [DOI] [PubMed] [Google Scholar]
  11. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  12. Lin Y. L., Blaschek H. P. Transformation of Heat-Treated Clostridium acetobutylicum Protoplasts with pUB110 Plasmid DNA. Appl Environ Microbiol. 1984 Oct;48(4):737–742. doi: 10.1128/aem.48.4.737-742.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Palosaari N. R., Rogers P. Purification and properties of the inducible coenzyme A-linked butyraldehyde dehydrogenase from Clostridium acetobutylicum. J Bacteriol. 1988 Jul;170(7):2971–2976. doi: 10.1128/jb.170.7.2971-2976.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Petersen D. J., Bennett G. N. Purification of acetoacetate decarboxylase from Clostridium acetobutylicum ATCC 824 and cloning of the acetoacetate decarboxylase gene in Escherichia coli. Appl Environ Microbiol. 1990 Nov;56(11):3491–3498. doi: 10.1128/aem.56.11.3491-3498.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Petersen D. J., Welch R. W., Rudolph F. B., Bennett G. N. Molecular cloning of an alcohol (butanol) dehydrogenase gene cluster from Clostridium acetobutylicum ATCC 824. J Bacteriol. 1991 Mar;173(5):1831–1834. doi: 10.1128/jb.173.5.1831-1834.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Young M., Minton N. P., Staudenbauer W. L. Recent advances in the genetics of the clostridia. FEMS Microbiol Rev. 1989 Dec;5(4):301–325. doi: 10.1111/j.1574-6968.1989.tb03402.x. [DOI] [PubMed] [Google Scholar]
  17. Youngleson J. S., Jones W. A., Jones D. T., Woods D. R. Molecular analysis and nucleotide sequence of the adh1 gene encoding an NADPH-dependent butanol dehydrogenase in the Gram-positive anaerobe Clostridium acetobutylicum. Gene. 1989 May 30;78(2):355–364. doi: 10.1016/0378-1119(89)90238-2. [DOI] [PubMed] [Google Scholar]
  18. Youngleson Jonathan S., Santangelo Joseph D., Jones David T., Woods David R. Cloning and Expression of a Clostridium acetobutylicum Alcohol Dehydrogenase Gene in Escherichia coli. Appl Environ Microbiol. 1988 Mar;54(3):676–682. doi: 10.1128/aem.54.3.676-682.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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