Use of recombination techniques to examine the structure of the csg locus of Myxococcus xanthus (original) (raw)
Summary
The myxobacteria are among the simplest organisms with a developmental cycle that is dependent on cell cooperation, and they provide an outstanding system with which to study genes involved in cell interactions. Myxococcus xanthus cells which acquire a csg mutation (formerly known as spoC) lose three different traits, the ability to sporulate, the ability to stimulate adjacent Csg cells to sporulate, and the ability to ripple. The boundaries of the csg locus were determined by transferring a recombinant DNA molecule containing all or part of the locus to Csg mutants and examining the sporulation and rippling phenotypes of the transductants. Three methods were used to integrate the csg locus into the chromosome. First, the entire molecule was integrated into the chromosome by a single homologous crossover. Second, a portion of the molecule was integrated into the chromosome by two flanking homologous crossovers. Third, the entire molecule was integrated into the chromosome by site-specific recombination at a bacteriophage attachment site. Together, these techniques suggested that all of the functions of the csg locus are carried on a DNA fragment of 1.9 kbp or less. The locus appears to contain two smaller units of function. Transposon insertions or deletions in the right end of the locus disrupted sporulation and intercellular complementation of Csg mutants for sporulation, but did not disrupt rippling. The intercellular complementation of Csg mutants may reflect a natural and necessary step in the sporulation of wild-type cells, since the ability to sporulate and the ability to stimulate Csg mutants to sporulate were inseparable by any of these methods.
Access this article
Subscribe and save
- Get 10 units per month
- Download Article/Chapter or eBook
- 1 Unit = 1 Article or 1 Chapter
- Cancel anytime Subscribe now
Buy Now
Price excludes VAT (USA)
Tax calculation will be finalised during checkout.
Instant access to the full article PDF.
Similar content being viewed by others
References
- Casadaban M, Cohen SN (1980) Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol 138:179–207
Google Scholar - Dworkin M, Kaiser D (1985) Cell interactions in myxobacterial growth and development. Science 230:18–24
Google Scholar - Feinberg AP, Vogelstein B (1983) A technique for radiolabelling DNA restriction endonuclease fragments to a high specific activity. Anal Biochem 132:6–13
Google Scholar - Grunstein M, Hogness D (1975) Colony hybridization: a method for isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci USA 72:3961–3965
Google Scholar - Hagen DC, Bretscher AP, Kaiser D (1978) Synergism between morphogenetic mutants of Myxococus xanthus. Dev Biol 64:284–296
Google Scholar - Hodgkin J, Kaiser D (1977) Cell-to-cell stimulation of motility in nonmotile mutants of Myxococcus. Proc Natl Acad Sci USA 74:2938–2942
Google Scholar - Kroos L, Kaiser D (1984) Construction of Tn_5 lac_, a transposon that fuses lacZ expression to endogenous promoters, and its introduction into Myxococcus xanthus. Proc Natl Acad Sci USA 81:5816–5820
Google Scholar - Kroos L, Kuspa A, Kaiser D (1986) A global analysis of developmentally regulated genes in Myxococcus xanthus. Dev Biol 117:252–266
Google Scholar - Jorgensen RA, Rothstein SJ, Reznikoff WS (1979) A restriction enzyme cleavage map of Tn_5_ and location of a region encoding neomycin resistance. Mol Gen Genet 177:65–72
Google Scholar - Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York
Google Scholar - O'Conner KA, Zusman DR (1983) Coliphage P1 mediated transfer of DNA from Escherichia coli to Myxococcus xanthus. Use for complementation and recombinational analysis. J Bacteriol 155:317–329
Google Scholar - Orndorff PE, Stellwag E, Starich T, Dworkin M, Zissler J (1983) Genetic and physical characterization of lysogeny by bacteriophage Mx8 in Myxococcus xanthus. J Bacteriol 154:772–779
Google Scholar - Reichenbach H (1965) Rhythmische Vorgänge bei der Schwarmentwicklung von Myxobakterien. Berl Dtsch Bot Ges 78:102–105
Google Scholar - Reichenbach H, Heunert HH, Kuczka H (1965) Schwarmentwicklung und Morphogenese bei Myxobakterien-Archangium, Myxococcus, Chondrococcus, Chondromyces. Film C893, Inst F Wissen, Film. Gottingen, Germany
Google Scholar - Shimkets LJ (1987) Control of morphogenesis in myxobacteria. CRC Crit Rev Microbiol 14:195–227
Google Scholar - Shimkets LJ, Kaiser S (1982a) Induction of coordinated movement of Myxococcus xanthus cells. J Bacteriol 152:451–461
Google Scholar - Shimkets LJ, Kaiser D (1982b) Murien components rescue developmental sporulation of Myxococcus xanthus. J Bacteriol 152:462–470
Google Scholar - Shimkets LJ, Gill RE, Kaiser D (1983) Developmental cell interactions in Myxococcus xanthus and the spoC locus. Proc Natl Acad Sci USA 80:1406–1410
Google Scholar - Stellwag E, Fink JM, Zissler J (1985) Physical characterization of the genome of the Myxococcus xanthus bacteriophage Mx8. Mol Gen Genet 199:123–132
Google Scholar - Sternberg N, Hoess R (1983) The molecular genetics of bacteriophage P1. Annu Rev Genet 17:123–154
Google Scholar - Youngman P, Zuber P, Perkins JB, Sandman K, Igo M, Losick R (1985) New ways to study developmental genes in sporeforming bacteria. Science 228:285–291
Google Scholar
Author information
Authors and Affiliations
- Department of Microbiology, University of Georgia, 30602, Athens, GA, USA
Lawrence J. Shimkets & Sheilah J. Asher
Authors
- Lawrence J. Shimkets
You can also search for this author inPubMed Google Scholar - Sheilah J. Asher
You can also search for this author inPubMed Google Scholar
Additional information
Communicated by N.D.I. Grindley
Rights and permissions
About this article
Cite this article
Shimkets, L.J., Asher, S.J. Use of recombination techniques to examine the structure of the csg locus of Myxococcus xanthus.Mol Gen Genet 211, 63–71 (1988). https://doi.org/10.1007/BF00338394
- Received: 20 August 1987
- Issue Date: January 1988
- DOI: https://doi.org/10.1007/BF00338394