Abstract
Under conditions where plasmid-carriage is deleterious to the cell, evolutionary changes may be expected which result in an attenuation of the deleterious effect of the plasmid. During long-term growth in glucose-limited continuous culture, initiated with a single clone of Escherichia coli containing a derivative of the plasmid pBR322, a structural change arose in the plasmid and predominated in the plasmid-containing sector of the population. This variant possessed a 2.25 kb deletion encompassing the tetracycline resistance operon as well as a region of about 1.5 kb upstream from this operon. Competition experiments involving strains carrying the plasmid with the spontancous deletion, and strains carrying plasmids with artificially constructed deletions, revealed that deletion of this region of the plasmid, involving loss of tetracycline resistance, resulted in an increment in fitness of between 10 and 20%. From the magnitude of the growth advantage, we conclude that the attenuation of the deleterious effect of the plasmid was mainly due to a reduction in the plasmid mediated interference in the metabolism of the cell caused by a deletion of the tetracycline resistance gene.
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Anderson, R. P. & Roth, J. R., 1977. Tandem genetic duplications in phage and bacteria. Ann. Rev. Microbiol. 31: 473–505.
Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. & Struhl, K., 1989. Current Protocols in Molecular Biology, Vol. 1. Greene publishing Associates and Wiley-Interscience.
Boe, L., Gerdes, K. & Molin, S., 1987. Effects of genes exerting growth inhibition and plasmid stability on plasmid maintenance. J. Bacteriol. 169: 4646–4650.
Bouma, J. E. & Lenski, R. E., 1988. Evolution of a bacteria/plasmid association. Nature 335: 351–352.
Boyko, W. L. & Ganschow, R. E., 1982. Rapid identification of Escherichia coli transformed by pBR322 carrying inserts at the PstI site. Anal. Biochem. 122: 885–888.
Chao, L. & Bremer, H., 1986a. Effect of the bacterial growth rate on replication control of plasmid pBR322 in Escherichia coli. Mol. Gen. Genet. 203: 143–149.
Chao, L. & Bremer, H., 1986b. Effect of relA function on the replication of plasmid pBR322 in Escherichia coli. Gen. Genet. 203: 150–153.
Cheah, U. E., Weigand, W. A. & Stark, B. C., 1987. Effects of recombinant plasmid size on cellular processes in Escherichia coli. Plasmid 18: 127–134.
Chiang, C. S. & Bremer, H., 1988. Stability of pBR322-derived plasmids. Plasmid 20: 207–220.
Eckert, B. & Beck, C. F., 1989. Overproduction of transposon Tn10-encoded resistance protein results in cell death and loss of membrane potential. J. Bacteriol. 171: 3557–3559.
Edlund, T. & Normark, S., 1981. Recombination between short DNA homologies causes tandem duplication. Natur 292: 269–271.
Engberg, B. & Nordstrom, K., 1975. Replication of R-factor R1 in Escherichia coli K12 at different growth rates. J. Bacteriol. 123: 179–186.
Godwin, D. & Slater, J. H., 1979. The influence of the growth environment on the stability of a drug resistance plasmid in Escherichia coli K12. J. Gen. Microbiol. 111: 201–210.
Heffron, F., Kostriken, R., Morita, C. & Parker, R., 1981. Tn3 encodes a site-specific recombination system: identification of essential sequences, genes, and the actual site of recombination. Cold Spring Harbor Symp. Quant. Biol. 45: 259–268.
Helling, R. B., Kinney, T. & Adams, J., 1981. The maintenance of plasmid-containing organisms in populations of Escherichia coli. J. Gen. Microbiol. 123: 129–141.
Helling, R. B., Vargas, C. N. & Adams, J., 1987. Evolution of Escherichia coli during growth in a constant environment. Genetics 116: 349–358.
Horiuchi, T., Horiuchi, S. & Novick, A., 1963. The genetic basis of hypersynthesis of β-galactosidase. Genetics 48: 157–169.
Hwu, H. R., Roberts, J. W., Davidson, E. H. & Britten, R. J., 1986. Insertion and/or deletion of many repeated DNA sequences in human and higher ape evolution. Proc. Nat. Acad. Sci. USA 83: 3875–3879.
Inselburg, J., 1978. ColE1 plasmid mutants affecting growth of an Escherichia coli recB recC sbcB mutant. J. Bacteriol. 133: 433–436.
Jessop, A. P. & Clugston, C., 1985. Amplification of the ArgF region in strain HfrP4X of E. coli K12. Mol. Gen. Genet. 201: 347–350.
Jones, I. M., Primrose, S. B., Robinson, A. & Ellwood, D. C., 1980. Maintenance of some ColE1-type plasmids in continuous culture. Mol. Gen. Genet. 180: 579–584.
Kubitschek, H. E., 1970. Introduction to Research with Continuous Cultures. Prentice-Hall Inc., Englewood Cliffs, New Jersey.
Lee, S. W. & Edlin, G., 1985. Expression of tetracycline resistance in pBR322 derivatives reduces the reproductive fitness of plasmid-containing E. coli. Gene 39: 173–180.
Lee, S. C., Gerding, D. N. & Cleary, P. P., 1984. Plasmid macro-evolution in a nosocomial environment; demonstration of a persistent molecular polymorphism and construction of a cladistic phylogeny on the basis of restriction data. Mol. Gen. Genet. 194: 173–178.
Levin, B. R. & Lenski, R. E., 1983. Coevolution in bacteria and their viruses and plasmids, pp. 99–127. In. Coevolution. Edited by D. J. Futuyma and M. Slatkin. Sinauer Assoc., Sunderland, Mass.
Lundquist, P. D. & Levin, B. R., 1986. Transitory depression and the maintenance of conjugative plasmids. Genetics 113: 483–497.
Maniatis, T., Fritsch, E. F. & Sambrook, J., 1982. Molecular Cloning-A Laboratory Manual. Cold Spring Harbor Laboratories. Cold Spring Harbor, New York.
Melling, J., Ellwood, D. C. & Robinson, A., 1977. Survival of R-factor carrying Escherichia coli in mixed cultures in the chemostat. FEMS Microbiol. Lett. 2: 87–89.
Miller, R. D., Dykhuizen, D. E., Green, L. & Hartl, D. L., 1984. Specific deletion occurring in the directed evolution of 6-phosphogluconate dehydrogenase in Escherichia coli. Genetics 108: 765–772.
Modi, R. I. & Adams, J., 1991. Coevolution in bacterial-plasmid populations. Evolution 45: 656–667.
Nguyen, T. N. M., Phan, Q. G., Duong, L. P., Bertrand, K. P. & Lenski, R. E., 1989. Effects of carriage and expression of the Tn10 tetracycline-resistance operon on the fitness of E. coli K12. Mol. Biol. Evol. 6: 213–225.
Noack, D., Roth, M., Guether, R., Muller, G., Undisz, K., Hoffmeier, C. & Gaspar, S., 1981. Maintenance and genetic stability of vector plasmids pBR322 and pBR325 in Escherichia coli K12 strains grown in a chemostat. Mol. Gen. Genet. 184: 121–124.
Peden, K. W. C., 1983. Revised sequence of the tetracycline-resistance gene of pBR322. Gene 22: 277–280.
Rigby, P. W. J., Burleigh, B. D. & Hartley, B. S., 1974. Gene duplication in experimental enzyme evolution. Nature 251: 200–204.
Roth, J. R. & Schmid, M., 1981. Arrangement and rearrangement of the bacterial genome. Stadler symp. 13: 53–70.
Roth, M., Muller, G. & Noack, D., 1980. Loss of the multi-copy resistance plasmid pBR325 from Escherichia coli CY2354 pBR325 during continuous cultivation, pp. 143–146. In. Fourth International Symposium on Antibiotic Resistance. Edited by S. Mitsuhasi, L. Rosival and V. Krcmery. Aviceneum Prague and Springer-Verlag West, Berlin.
Sonti, R. V. & Roth, J. R., 1989. Role of gene duplication in the adaptation of Salmonella typhimurium to growth on limiting carbon sources. Genetics 123: 19–28.
Tait, R. C. & Boyer, H. W., 1978. On the nature of tetracycline resistance controlled by the plasmid pSC101. Cell 13: 73–81.
Taxis Du Poet, P., Arcand, Y., Bernier, R. Jr., Barbotin, J.-N. & Thomas, D., 1987. Plasmid stability in immobilized and free recombinant E. coli JM105(pKK223–200): Importance of oxygen diffusion, growth rate, and plasmid copy number. Appl. Environ. Microbiol. 53: 1548–1555.
Tlsty, D. T., Albertini, A. M. & Miller, J. H., 1984. Gene amplification in the lac region of E. coli. Cell 37: 217–224.
Warnes, A. & Stephenson, J. R., 1986. The insertion of large pieces of foreign genetic material reduces the stability of bacterial plasmids. Plasmid 16: 116–123.
Weinberger, M. & Helmstetter, C. E., 1979. Chromosome replication and cell division in plasmid-containing E. coli B/R. J. Bacteriology 137: 1151–1157.
Wouters, J. T. M., Driehuis, F. L., Polaczek, P. J., van Oppenraay, M.-L.H.A. & van Andel, J. G., 1980. Persistence of the pBR322 plasmid in Escherichia coli K12 grown in chemostat cultures. Antonie van Leeuwenhoek. Jour. Microbiol. Serol. 45: 353–362.
Wouters, J. T. M. & van Andel, J. G., 1979. R-plasmid persistance in Escherichia coli grown in chemostat cultures. Antonie van Leeuwenhoek. Jour. Microbiol. Serol. 45: 317–318.
Zund, P. & Lebek, G., 1980. Generation time-prolonging R plasmids: Correlation between increases in the generation time of Escherichia coli caused by R-plasmids and their molecular size. Plasmid 3: 65–69.
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Modi, R.I., Wilke, C.M., Rosenzweig, R.F. et al. Plasmid macro-evolution: selection of deletions during adaptation in a nutrient-limited environment. Genetica 84, 195–202 (1991). https://doi.org/10.1007/BF00127247
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DOI: https://doi.org/10.1007/BF00127247