Construction of Genetically Engineered Microorganisms for Overexpression of xylE Gene Encoding Catechol 2,3-dioxygenase and the Functional Stability of the Recombinant Plasmid pSW3a Containing xylE in Aquatic Environment

  • Han, Hyo-Yung (Department of Biology, Sookmyung Womens University) ;
  • Kim, Chi-Kyung (Chungbuk National University) ;
  • Park, Yong-Keun (Research Center for Molecular Microbiology, Seoul National University) ;
  • Ka, Jong-Ok (Research Center for Molecular Microbiology, Seoul National University) ;
  • Lee, Byeong-Jae (Research Center for Molecular Microbiology, Seoul National University) ;
  • Min, Kyung-Hee (Department of Biology, Sookmyung Womens University)
  • 발행 : 1996.12.01

초록

The regulation of xylE gene expression was examined by using vector promoter and construction of genetically engineered microorganisms (GEMs) for application in microcosm. When the xylE gene wsa subcloned into pBluscript SK(+) under the control of lac promoter (pTY1) in E. coli, and the expression was induced by IPTG, the enzyme activity of catechol 2, 3-dioxygenase was increased 4.7 times more than that of the crude extracts from transformants harboring pTY1. We suggest that the xylE gene has its own promoter at the upstream portion, because it was able to be expressed even in the absence of IPTG. A recombinant plasmid, pSW3a harboring the xylE gene under the T7 promotor, showed the activity of 14.5 units/mg protein, higher than that of parental strain, E. coli PYT1. The xylE gene in recombinant plasmid pSW3a was used as reporter gene for the application in microcosm ecosystem, since it was used for detection of xylE-positive clones by catechol spray on the agar plates. The pSW3a in E. coli was introduced into Pseudomonas patida to construct GEM strain, and examined for the exxpression and functional stability in microcosms.

키워드

참고문헌

  1. Experimental microbial ecology Enumeration and estimation of biomass of microbial components in the biosphere Atlas, R.M.;R.G. Burns(ed.);J.H. Slater(ed.)
  2. Adv. Microb. Ecol. v.4 The immunofluorescence approach in microbial ecology Bohlool, B.B.;E.L. Schmidt
  3. Bull. Ecol. Soc. Am. v.65 Report on workshop on possible ecological and evolutionary impacts of bioengineered organisms released into the environment Brown, T.H.;R.K. Colwell;R.E. Lenski;B.R. Levin;M. Lloyd;P.J. REgal;D. Simberloff
  4. Proc. Natl. Acad. Sci. USA v.77 Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti Ditta, G.;S. Stanffield;D. Corbin;D. R. Helinski
  5. Appl. Environ. Microbiol. v.59 no.9 Production of L-dihydroxypheylalanine in Escherichia coli with the tyrosine phenol-lyase gene cloned from Erwinia herbicola Foor F.;N. Morin;K.A. Bostian
  6. World J. Microbiol. Biotechnol. v.7 Catechol-2,3-oxygenase production by genetically engineered Escherichia coli and its application to catechol determination Fujita, M.;T. Kamiya;M. Ike;Y. Kawagoshi;N. Shinohara
  7. Wat. Res. v.27 Accelerated phenol removal by amplifying the gene expression with a recombinant plasmid encoding catechol-2,3-oxygen ase Fujita, M.;M. Ike;T. Kamiya
  8. J. Ferment. Bioeng. v.79 Trichloromethyl ene degradation by genetically engineered bacteria carrying cloned phenol catabolic genes Fujita, M.;M. Ike;J. Hioki;K. Kataoka;M. Takeo
  9. Agric. Biol. Chem. v.49 Construction of broad host range cloning vectors for gram-negative bacteria Fukuda, M.;M. Ike;Yano
  10. Appl. Environ. Microbiol. v.49 Transfer of plasmid pBR322 and pBR325 in wastewater from laboratory strains of Escherichia coli to bacteria indigenous to the waste disposal system Gealt, M.A.;M.D. Chai;K.B. Alpert;J.C. Boyer
  11. Appl. Environ. Microbiol. v.54 DNA probe method for the delection of specific microorganisms in the soil bacterial community Holben, W.E.;J.K. Jansson;B.K. Chelm;J.M. Tiedje
  12. J. Bacteriol. v.172 xylE functions as an efficient reporter gene in Streptomyces spp.: Use for the study of galP1, a catabolitecontrolled promoter Ingram C.;M. Brawner;P. Youngman;J. Westpheling
  13. J. Bacterid. v.148 Molecular cloning of gene xylS of the TOL plasmid: Evidence for positive regulation of the xylDEFG operons by xylS Inouye S.;A. Nakazawa;T. Nakazawa
  14. J. Bacteriol. v.155 Molecular cloning of regulatory gene xylR and operator-promoter regions of the xylABC and xylDEFG operons of the TOL plasmid Inouye S.;A. Nakazawa;T. Nakazawa
  15. Gene. v.29 Nucleotide sequence of the promoter region of the xylDEFG operons on TOL plasmid of Pseudomonas putida. Inouye S.;A. Nakazawa;T. Nakazawa
  16. Appl. Environ. Microbiol. v.54 Regulation of 2,4,5-trichlorophenoxyacetic acid and chlorophenol metabolism in Pseudomonas cepacia AC1100 Karns, J.S.;S. Duttagupta;A.M. Chakrabarty
  17. J. Bacteriol. v.163 TOL plasmid pWW15 contains two nonhomologous independently regulated C230 genes Keil. H.;M.R. Lobens;P.A. Williams
  18. Kor. J. Microbiol. v.32 no.6 Cloning and nucleotide sequence analysis of xylE encoding catechol 2,3-dioxygenase from Pseudomonas putida SU10 Kim Y.J.;B.J. Lee;K.H. Min
  19. J. Microbiol. v.34 Structural and functional stability of the gene recombinant plasmid pCU103 in different water environments Kim C.K.;M.J. Kwak;S.G. Lee
  20. J. Biochem. v.117 Overexpression of Pseudomonas putida catechol-2,3-dioxygenase with high specific activity by genetically engineered Escherichia coli Kobayashi T.;T. Ishida;K. Horiike;Y. Takahara;N. Numao;A. Nakazawa;T. Nakazawa;M. Nozaki
  21. Biol/Technology v.5 EPA developing methods to assess environmental release Levin, M.A.;R. Seidler;A.W. Bourquin;J.W. Fowle Ⅲ;T. Barkay
  22. J. Biol. Chem. v.193 Protein measurement with the folin phenol reagent Lowry, O.H.;N.J. Rosebrough;A.L. Farr
  23. Appl. Environ. Microbiol. v.57 Survival and catabolic activity of natural and genetically engineered bacteria in a laboratory-scale activated-sludge unit McClure, N.C.;J.C. Fry;A.J. Weightman
  24. Arch. Biochem. Biophys. v.332 no.2 Characterization of the gene encoding catechol 2,3-dioxygenase of Alcaligenes sp. KF711: Overexpression, enzyme purification, and nucleotide sequencing Moon J.H.;K.R. Min;C.K. Kim;K.H. Min;Y.S. Kim
  25. J. Bacteriol. v.117 Role of catechol and methly catechols as inducers of aromatic metabolism in Pseudomonas putida Murray K.;P.A. Williams
  26. J. Microbiol. Methods v.7 DNA extraction and purification from sediments Ogram, A.;G.S. Sayler;T. Barkay
  27. Appl. Environ. Microbiol. v.58 Survival and function of a genetically engineered Pseudomonas in aquatic sediment microcosms Pipke, R.;I. Wagner-Dobler;K. Timmis;D.F. Dwyer
  28. Microbiol. Rev. v.51 Survival strategies of bacteria in the natural environment Roszak, D.B.;R.R. Colwell
  29. Abst. Gen. Meet. Am. Soc. Microbiol. v.92 Expression of Plasmodium folciparum erythrocyte binding antigen peptide 4 in Salmonellae Rubin, F. A.;B.K.L. Sim;L.E. Lindler;F. Koster;B. Frain;R.L. Warren
  30. Molecular cloning: A laboratory manual Sambrook J.;E.F. Fritsch;T. Maniatis
  31. Appl. Environ. Microbiol. v.58 no.1 Use of bioluminescence for detection of genetically engineered microorganisms released into the environment Shaw, J.J.;F. Dane;D. Geiger;J. W. Kloepper
  32. Biotechnology v.1 A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in gram-negative bacteria Simon, R.;U. Priefer;A. Puhler
  33. Appl. Environ. Microbial. v.56 Degradation of 1,4-dichlorobenzene by a Psudomonasa sp. Spain, J.C.;S.F. Nishino
  34. Bacteria in nature, Vol. 2, Methods and special applications in bacterial ecology v.2 Quantitative physiochemical characterization of bacterial habiratats White, D.C.;J.S. poindexter(ed.);E.R. Leadbetter(ed.().