Abstract
Establishment of a system for manipulative genetics in phototrophic sulfur bacteria of the family Chromatiaceae has mainly been hampered by the lack of reliable methods for growth of these organisms on agar surfaces, techniques for streaking, growth on selective media, screening for antibiotic resistance markers, and most importantly by the lack of a system for DNA transfer. We, therefore, developed minimal and complex agar media forChromatium vinosum strain D (DSM 180T), a representative of the purple sulfur bacteria. Sensitivity ofC. vinosum towards a broad range of antibiotics was tested in liquid cultures and solidified media, allowing us to select appropriate antibiotic resistance markers. Furthermore, a system for conjugative transfer of IncP-mobilizable plasmids fromEscherichia coli toC. vinosum was established. Broad-host-range IncQ vectors were mobilized toC. vinosum with the aid of plasmid RP4 either present extrachromosomally or integrated in the chromosome ofE. coli S17-1. Conjugation efficiencies of up to 1 were observed. Agarose gel electrophoretic analysis showed that transconjugants contained the transferred plasmids in addition to the two detectable plasmids of wild-typeC. vinosum. All genetic markers tested (kanamycin, gentamicin, ampicillin, amikacin, tetracycline) were expressed inC. vinosum. Furthermore, high-frequency transfer of plasmid RP4 fromC. vinosum toE. coli and toRhodospirillum rubrum K100 was demonstrated.
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References
Bazaral M, Helinski DR (1968) Circular DNA forms of colicinogenic factors E1, E2 and E3 fromEscherichia coli. J Mol Biol 36:185–194
Bolivar F, Backman K (1979) Plasmids ofEscherichia coli as cloning vectors. Methods Enzymol 68:245–267
Colbeau A, Kovacs KL, Chabert J, Vignais PM (1994) Cloning and sequences of the structural (hupDHI) genes for hydrogenase biosynthesis inThiocapsa roseopersicina. Gene 140:25–31
Datta N, Hedges RW, Shaw EJ, Sykes RB, Richmond MH (1971) Properties of an R factor fromPseudomonas aeruginosa. J Bacteriol 108:1244–1249
Dolata MM, Van Beeumen JJ, Ambler RP, Meyer TE, Cusanovich MA (1993) Nucleotide sequence of the heme subunit of flavocytochromec from the purple phototrophic bacterium,Chromatium vinosum. A 2.6 kilobase pair DNA fragment contains two multiheme cytochromes, a flavoprotein, and a homolog of human ankyrin. J Biol Chem 268:14426–14431
Donohue TJ, Kaplan S (1991) Genetic techniques in Rhodospirillaceae. Methods Enzymol 204:459–485
Eckhardt T (1978) A rapid method for the identification of plasmid desoxyribonucleic acid in bacteria. Plasmid 1:584–588
Gaju N, Pavon V, Marin I, Esteve I, Guerrero R, Amils R (1995) Chromosome map of the phototrophic anoxygenic bacteriumChromatium vinosum. FEMS Microbiol Lett 126:241–248
Gerlitz M, Hrabak O, Schwab H (1990) Partitioning of broad-host-range plasmid RP4 is a complex system involving site-specific recombination. J Bacteriol 172:6194–6203
Heisig P (1993) High-level fluoroquinolone resistance in aSalmonella typhimurium isolate due to alterations in bothgyrA andgyrB genes. J Antimicrob Chemother 32:367–377
Heisig P, Wiedemann B (1991) Use of a broad-host-rangegyrA plasmid for genetic characterization of fluoroquinolone-resistant Gram-negative bacteria. Antimicrob Agents Chemother 35: 2031–2036
Hunter CN, Mann NH (1992) Genetic manipulation of photosynthetic prokaryotes. In: Mann NH, Carr NG (eds) Photosynthetic prokaryotes. Plenum Press, New York, pp 153–179
Irgens RL (1983) Thioacetamide as a source of hydrogen sulfide for colony growth of purple sulfur bacteria. Curr Microbiol 8: 183–186
Kämpf C, Pfennig N (1980) Capacity of Chromatiaceae for chemotrophic growth. Specific respiration rates ofThiocystis violacea andChromatium vinosum. Arch Microbiol 127:125–135
Kern M, Klipp W, Klemme JH (1994) Increased nitrogenase-dependent H2 photoproduction byhup mutants ofRhodospirillum rubrum. Appl Environ Microbiol 60:1768–1774
Klipp W, Masepohl B, Pühler A (1988) Identification and mapping of nitrogen fixation genes ofRhodobacter capsulatus: duplication of anifA-nifB region. J Bacteriol 170:693–699
Lang FS, Oesterhelt D (1989) Gene transfer system forRhodopseudomonas viridis. J Bacteriol 171:4425–4435
Madigan MT (1986)Chromatium tepidum sp. n., a thermophilic photosynthetic bacterium of the family Chromatiaceae. Int J Syst Bacteriol 36:222–227
Nogales B, Guerrero R, Esteve I (1994) Susceptibility of various purple and green sulfur bacteria to different antimicrobial agents. FEMS Microbiol Lett 123:37–42
Olsen J, Woese CR, Overbeek R (1994) The winds of (evolutionary) change: breathing new life into microbiology. J Bacteriol 176:1–6
Pansegrau W, Lanka E, Barth PT, Figurski DH, Guiney DG, Haas D, Helinski DR, Schwab H, Stanisich VA, Thomas CM (1994) Complete nucleotide sequence of Birmingham IncPα plasmids. Compilation and comparative analysis. J Mol Biol 239:623–663
Pfennig N (1989) Genus I.Chromatium Perty 1852, 174Al. In: Staley JT, Bryant MP, Pfennig N, Holt JG (eds) Bergey’s manual of systematic bacteriology, vol 3. Williams & Wilkins, Baltimore, pp 1639–1643
Pfennig N, Trüper HG (1971) Type and neotype strains of the species of phototrophic bacteria maintained in pure culture. Int J Syst Bacteriol 21:19–24
Pfennig N, Trüper HG (1991) The family Chromatiaceae. In: Balows A, Trüper HG, Dworkin M, Harder W, Schleifer KH (eds) The prokaryotes. Springer, Berlin Heidelberg New York, pp 3200–3221
Pommerich AM (1989) Antibiotika-Resistenz und Resistenzplasmide bei phototrophen Bakterien: Antibiotika-Resistenz und konjugativer Transfer von Resistenzplasmiden beiRhodospirillum rubrum. Diploma thesis, University of Bonn, Germany
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor
Simon R, Priefer U, Pühler A (1983) A broad-host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram-negative bacteria. Biotechnology 1:784–791
Sistrom WR (1977) Transfer of chromosomal genes mediated by plasmid R68.45 inRhodopseudomonas sphaeroides. J Bacteriol 131:526–532
Steinbüchel A, Hustede E, Liebergesell M, Pieper U, Timm A, Valentin H (1992) Molecular basis for biosynthesis and accumulation of polyhydroxyalkanoic acid in bacteria. FEMS Microbiol Rev 9:217–230
Suyama Y, Gibson J (1966) Satellite DNA in photosynthetic bacteria. Biochem Biophys Res Commun 24:549–553
Taylor DP, Cohen SN, Clark WG, Marrs BL (1983) Alignment of genetic and restriction maps of the photosynthesis region of theRhodopseudomonas capsulata chromosome by a conjugation-mediated marker rescue technique. J Bacteriol 154:580–590
Thomas CM, Smith CA (1987) Incompatibility group P plasmids: genetics, evolution and use in genetic manipulation. Annu Rev Microbiol 41:77–101
Tucker WT, Pemberton JM (1979) Conjugation and chromosome transfer inRhodopseudomonas sphaeroides mediated by W and P group plasmids. FEMS Microbiol Lett 5:173–176
Viale AM, Kobayashi H, Akazawa T (1989) Expressed genes for plant-type ribulose 1,5-bisphosphate carboxylase/oxygenase in the photosynthetic bacteriumChromatium vinosum, which possesses two complete sets of genes. J Bacteriol 171:2391–2400
Viale AM, Kobayashi H, Akazawa T, Henikoff S (1991)rbcR (correction ofrcbR), 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 bacteriumChromatium vinosum. J Bacteriol 173:5224–5229
Waugh DS, Pace NR (1990) Complementation of an RNaseP RNA (rnpB) gene deletion inEscherichia coli by homologous genes from distantly related species. J Bacteriol 172:6316–6312
Weaver PF, Wall JD, Gest H (1975) Characterization ofRhodopseudomonas capsulata. Arch Microbiol 105:207–216
Willison JC (1993) Biochemical genetics revisited: the use of mutants to study carbon and nitrogen metabolism in the photosynthetic bacteria. FEMS Microbiol Rev 104:1–38
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Dedicated to Prof. Norbert Pfennig on the occasion of his 70th birthday
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Pattaragulwanit, K., Dahl, C. Development of a genetic system for a purple sulfur bacterium: Conjugative plasmid transfer inChromatium vinosum . Arch. Microbiol. 164, 217–222 (1995). https://doi.org/10.1007/BF02529974
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DOI: https://doi.org/10.1007/BF02529974