Cloning and Characterization of the Paraquat Resistance-Related Genes from Ochrobactrum anthropi JW-2

Ochrobactrum anthropi JW-2 유래의 Paraquat 내성유전자 PqrA의 주변 유전자군 분석

  • Bae Eun-Kyung (Biotechnology Division, Korea Forest Research Institute) ;
  • Lee Hyo-Shin (Biotechnology Division, Korea Forest Research Institute) ;
  • Won Sung-Hye (Division of Animal Science and Technology, College of Agriculture and Life Science, Gyeongsang National University) ;
  • Lee Byung-Hyun (Division of Animal Science and Technology, College of Agriculture and Life Science, Gyeongsang National University)
  • 배은경 (국립산림과학원 생물공학과) ;
  • 이효신 (국립산림과학원 생물공학과) ;
  • 원성혜 (경상대학교 동물자원과학부) ;
  • 이병현 (경상대학교 동물자원과학부)
  • Published : 2006.03.01


A 4,971 bp chromosomal DNA fragment containing the pqrA, paraquat resistance gene, was cloned from Ochrobactrum anthropi JW-2, and the complete nucleotide sequence was determined. Nucleotide and deduced amino acid sequences of the fragment revealed the presence of 4 complete ORFs (orf2, pqrA, orf3, orf4) and two incomplete ORFs(orf1, orf5). Orf1, pqrA, orf4 and orf5 exists at the direct strand but orf2 and orf3 exists at the reverse complementary strand. Orf1 which of incomplete sequences without start codon shares homology with ATP binding region of the response regulator receiver. Orf2 shares high homology with members of the tetR family of transcriptional repressor which have a helix-turn-helix (H-T-H) motif. Therefore, the orf2 is predicted as a transcriptional repressor of pqrA and is designated as pqrR2. Orf3 shares high homology with the members of the lysR family acting as a transcriptional activator which have both of a H-T-H motif at the N-terminal region and substrate binding domain at the C-terminal region. Therefore, the orf3 is predicted as a transcriptional activator of pqrA and is designated as pqrR1. Orf4 shows homology with the periplasmic substrate-binding protein of amino acid ABC transporter. Orf5 which of incomplete sequences without stop codon revealed the homology with the permeases protein of amino acid ABC transporter.


Paraquat;paraquat resistance-related genes;Ochrobactrum anthropi JW-2


  1. Cho, Y. H., E. J. Kim, H. J. Chung, J. H Choi, K. F. Chater, B. E. Ahn, J. H. Shin, and J. H. Roe. 2003. The pqrAB operon is responsible for paraquat resistance in Streptomyces coelicolor. J. Bacteriol. 185: 6756-6763
  2. Comai L., D. Facciotti, W. R. Hiatt, G. Thompson, R. E. Rosi, and D. M. Stalker. 1985. Expression in plants of mutant aroA gene from Salmonella typhimurium confers tolerance to glyphosate. Nature 317: 741-744
  3. Hanahan, D. 1983. Studies on transformation of Escherichia coli with plasmids. J. Mol. Biol. 166: 557-580
  4. Kowallik, K. v., B. Stoebe, J. Schaffran, P. Kroth-Pancic, and U. Freier. 1995. The chloroplast genome of a chlorophyll a+c-containing alga, Odontella sinensis. Plant Mol. Biol. Rep. 13: 336-342
  5. Sanger, F., S. Nicklen, and A. R. Coulson. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463-5467
  6. Stalker, D. M., W. R. Hiatt, and L. Comai. 1985. A single amino acid substitution in the enzyme 5-enolpyruvylshikimate-3-phosphate synthase confers resistance to the herbicide glyphosate. J. BioI Chern. 260: 4724-4728
  7. Windhovel, D. and B. Bowine. 1991. Identification of cfxR, an activator gene of autotrophic CO2 fixation in Alcaligenes eutrophus. Mol. Micobiol. 5: 2695-2705
  8. Won, S. H., B. H. Lee, H. S. Lee, and J. K. Jo. 2001. An Ochrobactrum anthropi gene eonferring paraquat resistance to the heterologous host Escherichia coli. Biochem. Biophys. Res. Comm. 285: 885-890
  9. Murgia, I., D. Tarantino, C. Vannini, M. Bracale, S. Carravieri, and C. Soave. 2004. Arabidopsis thaliana plants overexpressing thylakoidal ascorbate peroxidase show increased resistance to Paraquat-induced photooxidative stress and to nitric oxide-induced cell death. Plant J. 38: 940-953
  10. Sambrook, J. and D.W. Russell. 2001. Molecular cloning: a laborotory manual, 3rd (eds.), p. 116. Cold spring Harbor Laboratory Press, Cold Spring Harbor, N.Y
  11. Preston, C. 1994. Resistance to photosystem I disrupting herbicide. p. 61-82. In S. B. Powles and A. M. Holtum (eds.), Herbicide Resistance in plants: Biology and Biochemistry. Lewis Publishers, Boca Raton, FL
  12. Santiviago, C. A., J. A. Fuentes, S. M. Bueno, A. N. Trombert, A. A. Hildago, L. T. Socias, P. Youderian, and G. C. Mora. 2002. The Salmonella enterica sv. Typhimurium smvA, yddG and ompD (porin) genes are required for the efficient efflux of methyl viologen. Mol. Microbiol. 46: 687-698
  13. Kim, S. E., S. Y. Kim, S. H. Ahn, and J. C. Chun. 2005. Determination of paraquat-resistant biotype on Conyza canadensis and the resistant mechanism. Kor. J. Pesticide Sci. 9: 88-96
  14. Muraoka, S., R. Okumura, N. Ogawa, T. Nonaka, K. Miyashita, and T. Senda. 2003. Crystal structure of a fulllength lysR-type transcriptional regulator, CbnR: Unusual combination of two subunit forms and molecular bases for causing and changing DNA bend. J. Mol. Biol. 328: 555-566
  15. Schumacher, M. A., M. C. Miller, S. Grkovic, M. H. Brown, R. A. Skurray, and R. G. Brennan. 2001. Structural mechanisms of QacR induction and multidrug recognition. Science 294: 2158-2163
  16. Jo, J. K., S. H. Won, D. Y. Son, and B. H. Lee. 2004. Paraquat resistance of transgenic tobacco plants overexpressing the Ochrobactrum anthorpi pqrA gene. Biotechnol. Lett. 26: 1391-1396
  17. Stragier, P., O. Danos, and J. C. Patte. 1983. Regulation of diaminopimelate decarboxylase synthesis in Escherichia Coli. II. Nucleotide sequence of the lysA gene and its regulatory region. J. Mol. Biol. 168: 321-331
  18. Viale, A. M., H. Kobayashi, T. Akazawa, and S. Henikoff. 1991. RbcR [correction of rcbR], a gene coding for a member of the LysR family of transcriptional regulators, is located upstream of the expressed set of ribulose 1,5-bisphosphate carboxylase/oxygenase genes in the photosynthetic bacterium Chrornatiurn vinosurn. J. Bacteriol. 173: 5224-5229
  19. Won, S. H., B. H. Lee, and J. K. Jo. 2000. Characterization of a paraquat resistance of Ochrobactrum anthroi JW-2. Kor. J. Appl. Microbiol. Biotechnol. 28: 1-7
  20. Kerppola, R. E. and G. Ferro-Luzzi Ames. 1992. Topology of the hydrophobic membrane-bound components of the histidine preiplasmic permease. J. Biol. Chem. 267: 2329-2336
  21. Wissenbach, D., S. Six, J. Bongaerts, D. Ternes, S. Steinwachs, and G. Dnden. 1995. A third periplasmic transport system for L-arginine in Eschericia coli: molecular characterization of the artPIQMJ genes, arginine binding and transport. Mol. Microbiol. 17: 675-686
  22. Yu, T., Y. S. Li, X. F. Chen, J. H. Hu, X. C. Chang, and Y. G. Zhu. 2003. Transgenic tobacco plants overexpressing cotton glutathione S-transferase (GST) show enhanced resistance to methyl viologen. J. Plant Physiol. 160: 1305-131l
  23. Ye, B. and J. Cressel. 2000. Transient, oxidant-induced antioxidant transcript and enzyme levels correlate with greater oxidant-resistance in paraquat-resistant Conyza bonariensis. Planta 211: 50-61
  24. Barry, G. F., M. L. Taylor, S. R. Padgette, K. H. Kolacz, L. E. Hallas, G. Della-Cioppa, and G. M. Kishore. 1994. Cloning and expression in Escherichia coli of the glyphosate-toaminomethyl phos-phonic acid degrading activity from Achromobacter sp. strain LBAA. Monsanto Technical Report MSLB 13245, St. Louis
  25. Stalker, D. M., K. E. McBride, and L. D. Malyj. 1988. Herbicide resistance in transgenic plants expressing a bacterial detoxification gene. Science 242: 419