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Stable transformation of Chromobacterium violaceum with a broad-host-range plasmid


Stable transformants of Chromobacterium violaceum were obtained by high-voltage electroporation with a 7-kilobase binary plasmid. The technique was reliable, reproducible, and simple, with efficiencies of 105 transformants/μg of plasmid DNA. The electrical conditions that resulted in the highest efficiencies were short pulse length (4.4–4.5 ms) and high voltage (12.5 kV/cm). The numbers of transformants were almost the same during the growth exponential phase (variation at optical density) and resulted in the highest efficiencies at DNA concentration of 250 pg/ml. Saturation appeared to begin at 4 μg/ml of DNA. This method of C. violaceum transformation should enhance the genetic and biotechnological research by providing a valuable, widely used procedure of introducing DNA into this bacterium.

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  1. Alvarez-Morales A, Betancourt-Alvarez M, Kaluza K, Hennecke H (1986) Activation of the Bradyrhizobium japonicum nifH and nifDK operons is dependent on promoter-upstream DNA sequences. Nucleic Acids Res 27:4207–4227

  2. Andrighetti-Frohner R, Creczynski-Pasa A, Barardi C, Simoes C (2003) Cytotoxicity and potential antiviral evaluation of violacein produced by Chromobacterium violaceum. Mem Inst Oswaldo Cruz 98:843–848

  3. Brazilian National Genome Project Consortium (2003) The complete genome sequence of Chromobacterium violaceum reveals remarkable and exploitable bacterial adaptability. Proc Natl Acad Sci U S A 100:11660–11665

  4. Chattopadhyay A, Kumar V, Bhat N, Rao PLNG (2002) Chromobacterium violaceum Infection: a rare but frequently fatal disease. J Pediatr Surg 37:108–110

  5. Covert SF, Kapoor P, Lee M-H, Briley A, Nairn CJ (2001) Agrobacterium tumefaciens-mediated transformation of Fusarium circinatum. Mycol Res 105:259–264

  6. Ditta G, Stanfield S, Corbin D, Helinski DR (1980) Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A 77:7347–7351

  7. Dower WJ, Miller JF, Ragsdale CW (1988) High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res 16:6127–6145

  8. Duran N, Menck CF (2001) Chromobacterium violaceum: a review of pharmacological and industrial perspectives. Crit Rev Microbiol 27:201–222

  9. Hattermann DR, Stacey G (1990) Efficient DNA transformation of Bradyrhizobium japonicum by electroporation. Appl Environ Microbiol 56:833–836

  10. Iwai M, Katoh H, Katayama M, Ikeuchi M (2004) Improved genetic transformation of the thermophilic cyanobacterium, Thermosynechococcus elongatus BP-1. Plant Cell Physiol 45:171–175

  11. Jacobs MA, Alwood A, Thaipisuttikul I, Spencer D, Haugen E, Ernst S, Will O, Kaul R, Raymond C, Levy R, Chun-Rong L, Guenthner D, Bovee D, Olson MV, Manoil C (2003) Comprehensive transposon mutant library of Pseudomonas aeruginosa. Proc Natl Acad Sci U S A 100:14339–14344

  12. Koide T, Silva Neto JF, Gomes SL, Marques MV (2004) Insertional transposon mutagenesis in the Xylella fastidiosa citrus variegated chlorosis strain with transposome. Curr Microbiol 48:247–250

  13. Lee MD, Odom JM, Buchanan RJ Jr (1998) New perspectives on microbial dehalogenation of chlorinated solvents: insights from the field. Annu Rev Microbiol 52:423–452

  14. Leon LL, Miranda CC, De Souza AO, Duran N (2001) Antileishmanial activity of the violacein extracted from Chromobacterium violaceum. J Antimicrob Chemother 48:449–450

  15. Martinez R, Velludo MASL, Santos VR, Dinamarco PV (2000) Chromobacterium violaceum infection in Brazil. A case report. Rev Inst Med Trop Sao Paulo 42:111–113

  16. Matz C, Deines P, Boenigk J, Arndt H, Eberl L, Kjelleberg S, Jurgens K (2004) Impact of violacein-producing bacteria on survival and feeding of bacterivorous nanoflagellates. Appl Environ Microbiol 70:1593–1599

  17. Miller JF, Dower WJ, Tompkins LS (1988) High voltage electroporation of bacteria: genetic transformation of Campylobacter jejuni with plasmid DNA. Proc Natl Acad Sci U S A 85:856–860

  18. Monteiro PB, Teixeira DC, Palma RR, Garnier M, Bove JM, Renaudin J (2001) Stable transformation of the Xylella fastidiosa citrus variegated chlorosis strain with oriC plasmids. Appl Environ Microbiol 67:2263–2269

  19. Moss MO, Ryall C (1981) The genus Chromobacterium. In: Starr M, Stolp H, Truper H, Balows A, Schlegel H (eds) The prokaryotes. Springer, Berlin Heidelberg New York, pp 1355–1363

  20. Onai K, Morishita M, Kaneko T, Tabata S, Ishiura M (2004) Natural transformation of the thermophilic cyanobacterium Thermosynechococcus elongatusBP-1: a simple and efficient method for gene transfer. Mol Genet Genomics 271:50–59

  21. Sambrook J, Russel DW (2001) Molecular cloning: A laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

  22. Steinbuchel A, Hein S (2001) Biochemical and molecular basis of microbial synthesis of polyhydroxyalkanoates in microorganisms. Adv Biochem Eng Biotechnol 71:81–123

  23. Stephens C (2004) Microbial genomics: Tropical treasure? Curr Biol 14:R65–R66

  24. Zinn M, Witholt B, Egli T (2001) Occurrence, synthesis and medical application of bacterial polyhydroxyalkanoate. Adv Drug Deliv Rev 53:5–21

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We thank S.C. da Silva and A. Schrank for critically reading this manuscript. This work was supported by grants from Brazilian National Research Council-CNPq/MCT-Proc 680220/00-5 and Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS). L.B, R.C., and S.W. are recipients of scholarships from CNPq (Functional Genome) and F.H.S. from FAPERGS.

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Correspondence to I. S. Schrank.

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Broetto, L., Cecagno, R., Sant'Anna, F.H. et al. Stable transformation of Chromobacterium violaceum with a broad-host-range plasmid. Appl Microbiol Biotechnol 71, 450–454 (2006).

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  • Transformation Efficiency
  • Pulse Length
  • Violacein
  • High Transformation Efficiency
  • Chromobacterium Violaceum