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. 1995 Oct;177(20):5748–5755. doi: 10.1128/jb.177.20.5748-5755.1995

Cloning and expression of the s-triazine hydrolase gene (trzA) from Rhodococcus corallinus and development of Rhodococcus recombinant strains capable of dealkylating and dechlorinating the herbicide atrazine.

Z Q Shao 1, W Seffens 1, W Mulbry 1, R M Behki 1
PMCID: PMC177393  PMID: 7592318

Abstract

We used degenerate oligodeoxyribonucleotides derived from the N-terminal sequence of the s-triazine hydrolase from Rhodococcus corallinus NRRL B-15444R in an amplification reaction to isolate a DNA segment containing a 57-bp fragment from the trzA gene. By using the nucleotide sequence of this fragment, a nondegenerate oligodeoxyribonucleotide was synthesized and used to screen a genomic library of R. corallinus DNA for fragments containing trzA. A 5.3-kb PstI fragment containing trzA was cloned, and the nucleotide sequence of a 2,450-bp region containing trzA was determined. No trzA expression was detected in Escherichia coli or several other gram-negative bacteria. The trzA gene was subcloned into a Rhodococcus-E. coli shuttle vector, pBS305, and transformed into several Rhodococcus strains. Expression of trzA was demonstrated in all Rhodococcus transformants. Rhodococcus sp. strain TE1, which possesses the catabolic gene (atrA) for the N-dealkylation of the herbicides atrazine and simazine, was able to dechlorinate the dealkylated metabolites of atrazine and simazine when carrying the trzA gene on a plasmid. A plasmid carrying both atrA and trzA was constructed and transformed into three atrA- and trzA-deficient Rhodococcus strains. Both genes were expressed in the transformants. The s-triazine hydrolase activity of the recombinant strains carrying the trzA plasmid were compared with that of the R. corallinus strain from which it was derived.

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Selected References

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  1. 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]
  2. Eaton R. W., Karns J. S. Cloning and analysis of s-triazine catabolic genes from Pseudomonas sp. strain NRRLB-12227. J Bacteriol. 1991 Feb;173(3):1215–1222. doi: 10.1128/jb.173.3.1215-1222.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Eaton R. W., Karns J. S. Cloning and comparison of the DNA encoding ammelide aminohydrolase and cyanuric acid amidohydrolase from three s-triazine-degrading bacterial strains. J Bacteriol. 1991 Feb;173(3):1363–1366. doi: 10.1128/jb.173.3.1363-1366.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Mandelbaum R. T., Allan D. L., Wackett L. P. Isolation and Characterization of a Pseudomonas sp. That Mineralizes the s-Triazine Herbicide Atrazine. Appl Environ Microbiol. 1995 Apr;61(4):1451–1457. doi: 10.1128/aem.61.4.1451-1457.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Morales V. M., Bäckman A., Bagdasarian M. A series of wide-host-range low-copy-number vectors that allow direct screening for recombinants. Gene. 1991 Jan 2;97(1):39–47. doi: 10.1016/0378-1119(91)90007-x. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Mulbry W. W. The aryldialkylphosphatase-encoding gene adpB from Nocardia sp. strain B-1: cloning, sequencing and expression in Escherichia coli. Gene. 1992 Nov 2;121(1):149–153. doi: 10.1016/0378-1119(92)90174-n. [DOI] [PubMed] [Google Scholar]
  9. Radosevich M., Traina S. J., Hao Y. L., Tuovinen O. H. Degradation and mineralization of atrazine by a soil bacterial isolate. Appl Environ Microbiol. 1995 Jan;61(1):297–302. doi: 10.1128/aem.61.1.297-302.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. 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]
  13. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  14. Yanze-Kontchou C., Gschwind N. Mineralization of the herbicide atrazine as a carbon source by a Pseudomonas strain. Appl Environ Microbiol. 1994 Dec;60(12):4297–4302. doi: 10.1128/aem.60.12.4297-4302.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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