Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1993 Mar;175(5):1272–1277. doi: 10.1128/jb.175.5.1272-1277.1993

Evolution of bacteriophage T7 in a growing plaque.

J Yin 1
PMCID: PMC193211  PMID: 8444789

Abstract

The emergence of mutants during the 10(9)-fold amplification of a bacteriophage was spatially resolved in a growing plaque. When wild-type phage T7 was grown on an Escherichia coli host which expressed an essential early enzyme of the phage infection cycle, the T7 RNA polymerase, mutant phage relying on this enzyme appeared in 10(8) phage replications and outgrew the wild type. Spatial resolution of the selection process was achieved by analyzing stab samples taken along a plaque radius. Different mutants were selected at different rates along different radii of the plaque, based on host range and restriction patterns of the isolates. The mutants deleted up to 11% of their genomes, including the gene for their own RNA polymerase. They gained an advantage over the wild type by replicating more efficiently, as determined by one-step growth cultures.

Full text

PDF
1272

Images in this article

1273Image
on p.1273
1274Image
on p.1274
1274Image
on p.1274
1276Image
on p.1276

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bauer G. J., McCaskill J. S., Otten H. Traveling waves of in vitro evolving RNA. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7937–7941. doi: 10.1073/pnas.86.20.7937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Campbell J. L., Richardson C. C., Studier F. W. Genetic recombination and complementation between bacteriophage T7 and cloned fragments of T7 DNA. Proc Natl Acad Sci U S A. 1978 May;75(5):2276–2280. doi: 10.1073/pnas.75.5.2276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chamberlin M., McGrath J., Waskell L. New RNA polymerase from Escherichia coli infected with bacteriophage T7. Nature. 1970 Oct 17;228(5268):227–231. doi: 10.1038/228227a0. [DOI] [PubMed] [Google Scholar]
  4. Demerec M, Fano U. Bacteriophage-Resistant Mutants in Escherichia Coli. Genetics. 1945 Mar;30(2):119–136. doi: 10.1093/genetics/30.2.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dunn J. J., Studier F. W. Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements. J Mol Biol. 1983 Jun 5;166(4):477–535. doi: 10.1016/s0022-2836(83)80282-4. [DOI] [PubMed] [Google Scholar]
  7. Eigen M., Gardiner W., Schuster P., Winkler-Oswatitsch R. The origin of genetic information. Sci Am. 1981 Apr;244(4):88-92, 96, et passim. doi: 10.1038/scientificamerican0481-88. [DOI] [PubMed] [Google Scholar]
  8. Eigen M. Selforganization of matter and the evolution of biological macromolecules. Naturwissenschaften. 1971 Oct;58(10):465–523. doi: 10.1007/BF00623322. [DOI] [PubMed] [Google Scholar]
  9. Hausmann R. Bacteriophage T7 genetics. Curr Top Microbiol Immunol. 1976;75:77–110. doi: 10.1007/978-3-642-66530-1_3. [DOI] [PubMed] [Google Scholar]
  10. Helling R. B., Goodman H. M., Boyer H. W. Analysis of endonuclease R-EcoRI fragments of DNA from lambdoid bacteriophages and other viruses by agarose-gel electrophoresis. J Virol. 1974 Nov;14(5):1235–1244. doi: 10.1128/jvi.14.5.1235-1244.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hillis D. M., Bull J. J., White M. E., Badgett M. R., Molineux I. J. Experimental phylogenetics: generation of a known phylogeny. Science. 1992 Jan 31;255(5044):589–592. doi: 10.1126/science.1736360. [DOI] [PubMed] [Google Scholar]
  12. McDonell M. W., Simon M. N., Studier F. W. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. doi: 10.1016/s0022-2836(77)80102-2. [DOI] [PubMed] [Google Scholar]
  13. Moffatt B. A., Studier F. W. Entry of bacteriophage T7 DNA into the cell and escape from host restriction. J Bacteriol. 1988 May;170(5):2095–2105. doi: 10.1128/jb.170.5.2095-2105.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Parvin J. D., Moscona A., Pan W. T., Leider J. M., Palese P. Measurement of the mutation rates of animal viruses: influenza A virus and poliovirus type 1. J Virol. 1986 Aug;59(2):377–383. doi: 10.1128/jvi.59.2.377-383.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pierce J. C., Masker W. Genetic deletions between directly repeated sequences in bacteriophage T7. Mol Gen Genet. 1989 Jun;217(2-3):215–222. doi: 10.1007/BF02464884. [DOI] [PubMed] [Google Scholar]
  16. Rosenberg A. H., Simon M. N., Studier F. W., Roberts R. J. Survey and mapping of restriction endonuclease cleavage sites in bacteriophage T7 DNA. J Mol Biol. 1979 Dec 25;135(4):907–915. doi: 10.1016/0022-2836(79)90519-9. [DOI] [PubMed] [Google Scholar]
  17. Scearce L. M., Pierce J. C., McInroy B., Masker W. Deletion mutagenesis independent of recombination in bacteriophage T7. J Bacteriol. 1991 Jan;173(2):869–878. doi: 10.1128/jb.173.2.869-878.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Studier F. W., Dunn J. J. Organization and expression of bacteriophage T7 DNA. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 2):999–1007. doi: 10.1101/sqb.1983.047.01.114. [DOI] [PubMed] [Google Scholar]
  19. Studier F. W. Genetic analysis of non-essential bacteriophage T7 genes. J Mol Biol. 1973 Sep 15;79(2):227–236. doi: 10.1016/0022-2836(73)90002-8. [DOI] [PubMed] [Google Scholar]
  20. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  21. Studier F. W., Rosenberg A. H., Simon M. N., Dunn J. J. Genetic and physical mapping in the early region of bacteriophage T7 DNA. J Mol Biol. 1979 Dec 25;135(4):917–937. doi: 10.1016/0022-2836(79)90520-5. [DOI] [PubMed] [Google Scholar]
  22. Studier F. W. The genetics and physiology of bacteriophage T7. Virology. 1969 Nov;39(3):562–574. doi: 10.1016/0042-6822(69)90104-4. [DOI] [PubMed] [Google Scholar]
  23. Yin J. A quantifiable phenotype of viral propagation. Biochem Biophys Res Commun. 1991 Jan 31;174(2):1009–1014. doi: 10.1016/0006-291x(91)91519-i. [DOI] [PubMed] [Google Scholar]
  24. Yin J., McCaskill J. S. Replication of viruses in a growing plaque: a reaction-diffusion model. Biophys J. 1992 Jun;61(6):1540–1549. doi: 10.1016/S0006-3495(92)81958-6. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES