This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Liu, Z.
Right arrow Articles by Bonnefoy, V.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Liu, Z.
Right arrow Articles by Bonnefoy, V.

 Previous Article  |  Next Article 

Journal of Bacteriology, April 2000, p. 2269-2276, Vol. 182, No. 8
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Construction and Characterization of a recA Mutant of Thiobacillus ferrooxidans by Marker Exchange Mutagenesis

Zhenying Liu, Nicolas Guiliani, Corinne Appia-Ayme, Françoise Borne, Jeanine Ratouchniak, and Violaine Bonnefoy*

Laboratoire de Chimie Bactérienne, Institut de Biologie Structurale et de Microbiologie, C.N.R.S., 13402 Marseille Cedex 20, France

Received 22 November 1999/Accepted 21 January 2000

To construct Thiobacillus ferrooxidans mutants by marker exchange mutagenesis, a genetic transfer system is required. The transfer of broad-host-range plasmids belonging to the incompatibility groups IncQ (pKT240 and pJRD215), IncP (pJB3Km1), and IncW (pUFR034) from Escherichia coli to two private T. ferrooxidans strains (BRGM1 and Tf-49) and to two collection strains (ATCC 33020 and ATCC 19859) by conjugation was analyzed. To knock out the T. ferrooxidans recA gene, a mobilizable suicide plasmid carrying the ATCC 33020 recA gene disrupted by a kanamycin resistance gene was transferred from E. coli to T. ferrooxidans ATCC 33020 by conjugation under the best conditions determined. The two kanamycin-resistant clones, which have retained the kanamycin-resistant phenotype after growth for several generations in nonselective medium, were shown to have the kanamycin resistance gene inserted within the recA gene, indicating that the recA::Omega -Km mutated allele was transferred from the suicide plasmid to the chromosome by homologous recombination. These mutants exhibited a slightly reduced growth rate and an increased sensitivity to UV and gamma  irradiation compared to the wild-type strain. However, the T. ferrooxidans recA mutants are less sensitive to these physical DNA-damaging agents than the recA mutants described in other bacterial species, suggesting that RecA plays a minor role in DNA repair in T. ferrooxidans.

* Corresponding author. Mailing address: Laboratoire de Chimie Bactérienne, Institut de Biologie Structurale et de Microbiologie, C.N.R.S., 31 chemin Joseph Aiguier, 13402 Marseille Cedex 20, France. Phone: (33) 491 164 146. Fax: (33) 491 718 914. E-mail: bonnefoy{at}ibsm.cnrs-mrs.fr.

Journal of Bacteriology, April 2000, p. 2269-2276, Vol. 182, No. 8
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • van Zyl, L. J., van Munster, J. M., Rawlings, D. E. (2008). Construction of arsB and tetH Mutants of the Sulfur-Oxidizing Bacterium Acidithiobacillus caldus by Marker Exchange. Appl. Environ. Microbiol. 74: 5686-5694 [Abstract] [Full Text]  
  • Han, Y. W., Ikegami, A., Chung, P., Zhang, L., Deng, C. X. (2007). Sonoporation Is an Efficient Tool for Intracellular Fluorescent Dextran Delivery and One-Step Double-Crossover Mutant Construction in Fusobacterium nucleatum. Appl. Environ. Microbiol. 73: 3677-3683 [Abstract] [Full Text]  
  • Letain, T. E., Kane, S. R., Legler, T. C., Salazar, E. P., Agron, P. G., Beller, H. R. (2007). Development of a Genetic System for the Chemolithoautotrophic Bacterium Thiobacillus denitrificans. Appl. Environ. Microbiol. 73: 3265-3271 [Abstract] [Full Text]  
  • Rivas, M., Seeger, M., Jedlicki, E., Holmes, D. S. (2007). Second Acyl Homoserine Lactone Production System in the Extreme Acidophile Acidithiobacillus ferrooxidans. Appl. Environ. Microbiol. 73: 3225-3231 [Abstract] [Full Text]  
  • Quatrini, R., Lefimil, C., Holmes, D. S., Jedlicki, E. (2005). The ferric iron uptake regulator (Fur) from the extreme acidophile Acidithiobacillus ferrooxidans. Microbiology 151: 2005-2015 [Abstract] [Full Text]  
  • Bruscella, P., Cassagnaud, L., Ratouchniak, J., Brasseur, G., Lojou, E., Amils, R., Bonnefoy, V. (2005). The HiPIP from the acidophilic Acidithiobacillus ferrooxidans is correctly processed and translocated in Escherichia coli, in spite of the periplasm pH difference between these two micro-organisms. Microbiology 151: 1421-1431 [Abstract] [Full Text]  
  • Duquesne, K., Lebrun, S., Casiot, C., Bruneel, O., Personne, J.-C., Leblanc, M., Elbaz-Poulichet, F., Morin, G., Bonnefoy, V. (2003). Immobilization of Arsenite and Ferric Iron by Acidithiobacillus ferrooxidans and Its Relevance to Acid Mine Drainage. Appl. Environ. Microbiol. 69: 6165-6173 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»