Abstract
The degradation of the herbicides EPTC (S-ethyl dipropylthiocarbamate) and atrazine (2-chloro-4-ethyl-amino-6-isopropylamino-1,3,5-triazine) is associated with an indigenous plasmid in Rhodococcus sp. strain TE1. Plasmid DNA libraries of Rhodococcus sp. strain TE1 were constructed in a Rhodococcus-Escherichia coli shuttle vector, pBS305, and transferred into Rhodococcus sp. strain TE3, a derivative of Rhodococcus sp. strain TE1 lacking herbicide degradation activity, to select transformants capable of growing on EPTC as the sole source of carbon (EPTC+). Analysis of plasmids from the EPTC+ transformants indicated that the eptA gene, which codes for the enzyme required for EPTC degradation, residues on a 6.2-kb KpnI fragment. The cloned fragment also harbored the gene required for atrazine N dealkylation (atrA). The plasmid carrying the cloned fragment could be electroporated into a number of other Rhodococcus strains in which both eptA and atrA were fully expressed. No expression of the cloned genes was evident in E. coli strains. Subcloning of the 6.2-kb fragment to distinguish between EPTC- and atrazine-degrading genes was not successful.
Full Text
The Full Text of this article is available as a PDF (204.8 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Behki R., Topp E., Dick W., Germon P. Metabolism of the herbicide atrazine by Rhodococcus strains. Appl Environ Microbiol. 1993 Jun;59(6):1955–1959. doi: 10.1128/aem.59.6.1955-1959.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
- Denome S. A., Olson E. S., Young K. D. Identification and Cloning of Genes Involved in Specific Desulfurization of Dibenzothiophene by Rhodococcus sp. Strain IGTS8. Appl Environ Microbiol. 1993 Sep;59(9):2837–2843. doi: 10.1128/aem.59.9.2837-2843.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Desomer J., Dhaese P., Montagu M. V. Transformation of Rhodococcus fascians by High-Voltage Electroporation and Development of R. fascians Cloning Vectors. Appl Environ Microbiol. 1990 Sep;56(9):2818–2825. doi: 10.1128/aem.56.9.2818-2825.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Finnerty W. R. The biology and genetics of the genus Rhodococcus. Annu Rev Microbiol. 1992;46:193–218. doi: 10.1146/annurev.mi.46.100192.001205. [DOI] [PubMed] [Google Scholar]
- Hashimoto Y., Nishiyama M., Ikehata O., Horinouchi S., Beppu T. Cloning and characterization of an amidase gene from Rhodococcus species N-774 and its expression in Escherichia coli. Biochim Biophys Acta. 1991 Feb 16;1088(2):225–233. doi: 10.1016/0167-4781(91)90058-t. [DOI] [PubMed] [Google Scholar]
- Hashimoto Y., Nishiyama M., Yu F., Watanabe I., Horinouchi S., Beppu T. Development of a host-vector system in a Rhodococcus strain and its use for expression of the cloned nitrile hydratase gene cluster. J Gen Microbiol. 1992 May;138(5):1003–1010. doi: 10.1099/00221287-138-5-1003. [DOI] [PubMed] [Google Scholar]
- Hill R., Hart S., Illing N., Kirby R., Woods D. R. Cloning and expression of Rhodococcus genes encoding pigment production in Escherichia coli. J Gen Microbiol. 1989 Jun;135(6):1507–1513. doi: 10.1099/00221287-135-6-1507. [DOI] [PubMed] [Google Scholar]
- Horn J. M., Harayama S., Timmis K. N. DNA sequence determination of the TOL plasmid (pWWO) xylGFJ genes of Pseudomonas putida: implications for the evolution of aromatic catabolism. Mol Microbiol. 1991 Oct;5(10):2459–2474. doi: 10.1111/j.1365-2958.1991.tb02091.x. [DOI] [PubMed] [Google Scholar]
- Ikehata O., Nishiyama M., Horinouchi S., Beppu T. Primary structure of nitrile hydratase deduced from the nucleotide sequence of a Rhodococcus species and its expression in Escherichia coli. Eur J Biochem. 1989 May 15;181(3):563–570. doi: 10.1111/j.1432-1033.1989.tb14761.x. [DOI] [PubMed] [Google Scholar]
- Mulbry W. W. Purification and Characterization of an Inducible s-Triazine Hydrolase from Rhodococcus corallinus NRRL B-15444R. Appl Environ Microbiol. 1994 Feb;60(2):613–618. doi: 10.1128/aem.60.2.613-618.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Singer M. E., Finnerty W. R. Construction of an Escherichia coli-Rhodococcus shuttle vector and plasmid transformation in Rhodococcus spp. J Bacteriol. 1988 Feb;170(2):638–645. doi: 10.1128/jb.170.2.638-645.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sunairi M., Watanabe T., Oda H., Murooka H., Nakajima M. Characterization of the genome of the Rhodococcus rhodochrous bacteriophage NJL. Appl Environ Microbiol. 1993 Jan;59(1):97–100. doi: 10.1128/aem.59.1.97-100.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tam A. C., Behki R. M., Khan S. U. Isolation and characterization of an s-ethyl-N,N-dipropylthiocarbamate-degrading Arthrobacter strain and evidence for plasmid-associated s-ethyl-N,N-dipropylthiocarbamate degradation. Appl Environ Microbiol. 1987 May;53(5):1088–1093. doi: 10.1128/aem.53.5.1088-1093.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vogelstein B., Gillespie D. Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci U S A. 1979 Feb;76(2):615–619. doi: 10.1073/pnas.76.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Warhurst A. M., Fewson C. A. Biotransformations catalyzed by the genus Rhodococcus. Crit Rev Biotechnol. 1994;14(1):29–73. doi: 10.3109/07388559409079833. [DOI] [PubMed] [Google Scholar]
- van der Meer J. R., de Vos W. M., Harayama S., Zehnder A. J. Molecular mechanisms of genetic adaptation to xenobiotic compounds. Microbiol Rev. 1992 Dec;56(4):677–694. doi: 10.1128/mr.56.4.677-694.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]