Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1973 Feb;113(2):798–812. doi: 10.1128/jb.113.2.798-812.1973

Rapid Mapping of Conditional and Auxotrophic Mutations in Escherichia coli K-12

Brooks Low 1
PMCID: PMC285295  PMID: 4570607

Abstract

The approximate genetic map locations of auxotrophic and conditional lethal mutations of Escherichia coli can be rapidly determined with replica plating techniques. A set of patches of 15 streptomycin-sensitive (StrS) Hfr strains with points of origin distributed around the map is replica plated onto a recombinant-selective plate with a lawn of StrR cells which carry an unmapped mutation. The map interval defined by the Hfr points of origin which are closest to the mutant locus is seen by the presence or absence of heavy patches of recombinants produced by transfer of early wild-type genes from the Hfrs. An alternative method is to replicate patches of different mutant strains (100 per plate) onto Hfr lawns; in this case more than 1,000 different mutants can be mapped in a single experiment in a few days. In this way, many types of mutations with similar phenotypes can be grouped as to approximate location on the genetic map. For ordering mutations within groups, the same replica plating methods can be used to cross F-prime derivatives of mutants with other mutants of the same group. Relative merits of these and other mapping methods of E. coli are discussed.

Full text

PDF
802

Images in this article

801Image
on p.801
804Image
on p.804
806Image
on p.806

Selected References

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

  1. ADELBERG E. A., BURNS S. N. Genetic variation in the sex factor of Escherichia coli. J Bacteriol. 1960 Mar;79:321–330. doi: 10.1128/jb.79.3.321-330.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abelson J. N., Gefter M. L., Barnett L., Landy A., Russell R. L., Smith J. D. Mutant tyrosine transfer ribonucleic acids. J Mol Biol. 1970 Jan 14;47(1):15–28. doi: 10.1016/0022-2836(70)90398-0. [DOI] [PubMed] [Google Scholar]
  3. Anderson W. F., Gorini L., Breckenridge L. Role of ribosomes in streptomycin-activated suppression. Proc Natl Acad Sci U S A. 1965 Oct;54(4):1076–1083. doi: 10.1073/pnas.54.4.1076. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bastarrachea F., Tam E., González A. Dominance of streptomycin sensitivity over dependence in Escherichia coli K12 merodiploids. Genetics. 1969 Dec;63(4):759–774. doi: 10.1093/genetics/63.4.759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bastarrachea F., Willetts N. S. The elimination by acridine orange of F30 from recombination-deficient strains of Escherichia coli K12. Genetics. 1968 Jun;59(2):153–166. doi: 10.1093/genetics/59.2.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Beckwith J. R., Signer E. R. Transposition of the lac region of Escherichia coli. I. Inversion of the lac operon and transduction of lac by phi80. J Mol Biol. 1966 Aug;19(2):254–265. doi: 10.1016/s0022-2836(66)80003-7. [DOI] [PubMed] [Google Scholar]
  7. Berg C. M., Curtiss R., 3rd Transposition derivatives of an Hfr strain of Escherichia coli K-12. Genetics. 1967 Jul;56(3):503–525. doi: 10.1093/genetics/56.3.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Breckenridge L., Gorini L. The dominance of streptomycin sensitivity re-examined. Proc Natl Acad Sci U S A. 1969 Mar;62(3):979–985. doi: 10.1073/pnas.62.3.979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clark A. J. Toward a metabolic interpretation of genetic recombination of E. coli and its phages. Annu Rev Microbiol. 1971;25:437–464. doi: 10.1146/annurev.mi.25.100171.002253. [DOI] [PubMed] [Google Scholar]
  10. Clowes R. C., Moody E. E. Chromosomal transfer from "recombination-deficient" strains of Escherichia coli K-12. Genetics. 1966 Apr;53(4):717–726. doi: 10.1093/genetics/53.4.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Curtiss R., 3rd, Caro L. G., Allison D. P., Stallions D. R. Early stages of conjugation in Escherichia coli. J Bacteriol. 1969 Nov;100(2):1091–1104. doi: 10.1128/jb.100.2.1091-1104.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. De Haan P. G., Hoekstra W. P., Verhoef C., Felix H. S. Recombination in Escherichia coli. 3. Mapping by the gradient of transmission. Mutat Res. 1969 Nov-Dec;8(3):505–512. doi: 10.1016/0027-5107(69)90067-0. [DOI] [PubMed] [Google Scholar]
  13. Glansdorff N. Pseudoinversions in the chromosome of Escherichia coli K-12. Genetics. 1967 Jan;55(1):49–61. doi: 10.1093/genetics/55.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Godson G. N. Isolation and partial characterization of temperature-sensitive mutants in ten loci that affect cell membrane synthesis in Escherichia coli: isolation and genetic sorting. J Bacteriol. 1973 Feb;113(2):813–824. doi: 10.1128/jb.113.2.813-824.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Goldschmidt E. P., Cater M. S., Matney T. S., Butler M. A., Greene A. Genetic analysis of the histidine operon in Escherichia coli K12. Genetics. 1970 Oct;66(2):219–229. doi: 10.1093/genetics/66.2.219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gottesman S., Beckwith J. R. Directed transposition of the arabinose operon: a technique for the isolation of specialized transducing bacteriophages for any Escherichia coli gene. J Mol Biol. 1969 Aug 28;44(1):117–127. doi: 10.1016/0022-2836(69)90408-2. [DOI] [PubMed] [Google Scholar]
  17. Guest J. R., Yanofsky C. Relative orientation of gene, messenger and polypeptide chain. Nature. 1966 May 21;210(5038):799–802. doi: 10.1038/210799a0. [DOI] [PubMed] [Google Scholar]
  18. HAYES W. The kinetics of the mating process in Escherichia coli. J Gen Microbiol. 1957 Feb;16(1):97–119. doi: 10.1099/00221287-16-1-97. [DOI] [PubMed] [Google Scholar]
  19. Hirota Y. THE EFFECT OF ACRIDINE DYES ON MATING TYPE FACTORS IN ESCHERICHIA COLI. Proc Natl Acad Sci U S A. 1960 Jan;46(1):57–64. doi: 10.1073/pnas.46.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hong J. S., Ames B. N. Localized mutagenesis of any specific small region of the bacterial chromosome. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3158–3162. doi: 10.1073/pnas.68.12.3158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. ISHIBASHI M., SUGINO Y., HIROTA Y. CHROMOSOMAL LOCATION OF THYMINE AND ARGININE GENES IN ESCHERICHIA COLI AND AN F' INCORPORATING THEM. J Bacteriol. 1964 Mar;87:554–561. doi: 10.1128/jb.87.3.554-561.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ippen K., Shapiro J. A., Beckwith J. R. Transposition of the lac region to the gal region of the Escherichia coli chromosome: isolation of lambda-lac transducing bacteriophages. J Bacteriol. 1971 Oct;108(1):5–9. doi: 10.1128/jb.108.1.5-9.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Joset F., Wood T. H. Modification of conjugation in Escherichia coli K-12 by ultraviolet irradiation. Genetics. 1966 Feb;53(2):343–356. doi: 10.1093/genetics/53.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kahn P. L. Isolation of high-frequency recombining strains from Escherichia coli containing the V colicinogenic factor. J Bacteriol. 1968 Jul;96(1):205–214. doi: 10.1128/jb.96.1.205-214.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kaiser A. D., Masuda T. Evidence for a prophage excision gene in lambda. J Mol Biol. 1970 Feb 14;47(3):557–564. doi: 10.1016/0022-2836(70)90322-0. [DOI] [PubMed] [Google Scholar]
  26. LEDERBERG J., LEDERBERG E. M. Replica plating and indirect selection of bacterial mutants. J Bacteriol. 1952 Mar;63(3):399–406. doi: 10.1128/jb.63.3.399-406.1952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  28. LOW B., WOOD T. H. A QUICK AND EFFICIENT METHOD FOR INTERRUPTION OF BACTERIAL CONJUGATION. Genet Res. 1965 Jul;6:300–303. doi: 10.1017/s001667230000416x. [DOI] [PubMed] [Google Scholar]
  29. LURIA S. E., BURROUS J. W. Hybridization between Escherichia coli and Shigella. J Bacteriol. 1957 Oct;74(4):461–476. doi: 10.1128/jb.74.4.461-476.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lederberg J, Cavalli L L, Lederberg E M. Sex Compatibility in Escherichia Coli. Genetics. 1952 Nov;37(6):720–730. doi: 10.1093/genetics/37.6.720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Low B. Formation of merodiploids in matings with a class of Rec- recipient strains of Escherichia coli K12. Proc Natl Acad Sci U S A. 1968 May;60(1):160–167. doi: 10.1073/pnas.60.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Low B., Gates F., Goldstein T., Söll D. Isolation and partial characterization of temperature-sensitive Escherichia coli mutants with altered leucyl- and seryl-transfer ribonucleic acid synthetases. J Bacteriol. 1971 Nov;108(2):742–750. doi: 10.1128/jb.108.2.742-750.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Low B. Inversion of transfer modes and sex factor-chromosome interactions in conjugation in Escherichia coli. J Bacteriol. 1967 Jan;93(1):98–106. doi: 10.1128/jb.93.1.98-106.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Low B. Low recombination frequency for markers very near the origin in conjugation in E. coli. Genet Res. 1965 Nov;6(3):469–473. doi: 10.1017/s0016672300004341. [DOI] [PubMed] [Google Scholar]
  35. MAAS R., MAAS W. K. Introduction of a gene from Escherichia coli B into HFR and F-strains of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1962 Nov 15;48:1887–1893. doi: 10.1073/pnas.48.11.1887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. MONOD J., COHEN-BAZIRE G., COHN M. Sur la biosynthèse de la beta-galactosidase (lactase) chez Escherichia coli; la spécificité de l'induction. Biochim Biophys Acta. 1951 Nov;7(4):585–599. doi: 10.1016/0006-3002(51)90072-8. [DOI] [PubMed] [Google Scholar]
  37. Marsh N. J., Duggan D. E. Ordering of mutant sites in the isoleucine-valine genes of Escherichia coli by use of merogenotes derived from F 14 : a new procedure for fine-structure mapping. J Bacteriol. 1972 Feb;109(2):730–740. doi: 10.1128/jb.109.2.730-740.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Miller J. H., Reznikoff W. S., Silverstone A. E., Ippen K., Signer E. R., Beckwith J. R. Fusions of the lac and trp Regions of the Escherichia coli Chromosome. J Bacteriol. 1970 Dec;104(3):1273–1279. doi: 10.1128/jb.104.3.1273-1279.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Norkin L. C. Marker-specific effects in genetic recombination. J Mol Biol. 1970 Aug;51(3):633–655. doi: 10.1016/0022-2836(70)90013-6. [DOI] [PubMed] [Google Scholar]
  40. PITTARD J., ADELBERG E. A. GENE TRANSFER BY F' STRAINS OF ESCHERICHIA COLI K-12. II. INTERACTION BETWEEN F-MEROGENOTE AND CHROMOSOME DURING TRANSFER. J Bacteriol. 1963 Jun;85:1402–1408. doi: 10.1128/jb.85.6.1402-1408.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. PITTARD J., ADELBERG E. A. GENE TRANSFER BY F' STRAINS OF ESCHERICHIA COLI K-12. III. AN ANALYSIS OF THE RECOMBINATION EVENTS OCCURRING IN THE F' MALE AND IN THE ZYGOTES. Genetics. 1964 Jun;49:995–1007. doi: 10.1093/genetics/49.6.995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. PITTARD J., RAMAKRISHNAN T. GENE TRANSFER BY F' STRAINS OF ESCHERICHIA COLI. IV. EFFECT OF A CHROMOSOMAL DELETION ON CHROMOSOME TRANSFER. J Bacteriol. 1964 Aug;88:367–373. doi: 10.1128/jb.88.2.367-373.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Pittard J., Walker E. M. Conjugation in Escherichia coli: recombination events in terminal regions of transferred deoxyribonucleic acid. J Bacteriol. 1967 Nov;94(5):1656–1663. doi: 10.1128/jb.94.5.1656-1663.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Press R., Glansdorff N., Miner P., De Vries J., Kadner R., Maas W. K. Isolation of transducing particles of phi-80 bacteriophage that carry different regions of the Escherichia coli genome. Proc Natl Acad Sci U S A. 1971 Apr;68(4):795–798. doi: 10.1073/pnas.68.4.795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. RAMAKRISHNAN T., ADELBERG E. A. REGULATORY MECHANISMS IN THE BIOSYNTHESIS OF ISOLEUCINE AND VALINE. II. IDENTIFICATION OF TWO OPERATOR GENES. J Bacteriol. 1965 Mar;89:654–660. doi: 10.1128/jb.89.3.654-660.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. STACEY K. A., SIMSON E. IMPROVED METHOD FOR THE ISOLATION OF THYMINE-REQUIRING MUTANTS OF ESCHERICHIA COLI. J Bacteriol. 1965 Aug;90:554–555. doi: 10.1128/jb.90.2.554-555.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Shapiro J. A., Adhya S. L. The galactose operon of E. coli K-12. II. A deletion analysis of operon structure and polarity. Genetics. 1969 Jun;62(2):249–264. doi: 10.1093/genetics/62.2.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Shapiro J., Machattie L., Eron L., Ihler G., Ippen K., Beckwith J. Isolation of pure lac operon DNA. Nature. 1969 Nov 22;224(5221):768–774. doi: 10.1038/224768a0. [DOI] [PubMed] [Google Scholar]
  49. Shimada K., Weisberg R. A., Gottesman M. E. Prophage lambda at unusual chromosomal locations. I. Location of the secondary attachment sites and the properties of the lysogens. J Mol Biol. 1972 Feb 14;63(3):483–503. doi: 10.1016/0022-2836(72)90443-3. [DOI] [PubMed] [Google Scholar]
  50. Spudich J. A., Horn V., Yanofsky C. On the production of deletions in the chromosome of Escherichia coli. J Mol Biol. 1970 Oct 14;53(1):49–67. doi: 10.1016/0022-2836(70)90045-8. [DOI] [PubMed] [Google Scholar]
  51. Stadler J., Adelberg E. A. Temperature dependence of sex-factor maintenance in Escherichia coli K-12. J Bacteriol. 1972 Jan;109(1):447–449. doi: 10.1128/jb.109.1.447-449.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Sunshine M. G., Kelly B. Extent of host deletions associated with bacteriophage P2-mediated eduction. J Bacteriol. 1971 Nov;108(2):695–704. doi: 10.1128/jb.108.2.695-704.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. TAYLOR A. L., THOMAN M. S. THE GENETIC MAP OF ESCHERICHIA COLI K-12. Genetics. 1964 Oct;50:659–677. doi: 10.1093/genetics/50.4.659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Taylor A L, Adelberg E A. Linkage Analysis with Very High Frequency Males of Escherichia Coli. Genetics. 1960 Sep;45(9):1233–1243. doi: 10.1093/genetics/45.9.1233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Taylor A. L. Current linkage map of Escherichia coli. Bacteriol Rev. 1970 Jun;34(2):155–175. doi: 10.1128/br.34.2.155-175.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Verhoef C., de Haan P. G. Genetic recombination in Escherichia coli. I. Relation between linkage of unselected markers and map distance. Mutat Res. 1966 Apr;3(2):101–110. doi: 10.1016/0027-5107(66)90023-6. [DOI] [PubMed] [Google Scholar]
  57. Wann M., Mahajan S. K., Wood T. H. Genetic mapping in Escherichia coli K-12 by radiation-induced crossing-over. J Bacteriol. 1970 Sep;103(3):601–606. doi: 10.1128/jb.103.3.601-606.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Wilkins B. M. Chromosome transfer from F-lac+ strains of Escherichia coli K-12 mutant at recA, recB, or recC. J Bacteriol. 1969 May;98(2):599–604. doi: 10.1128/jb.98.2.599-604.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Wilkins B. M., Howard-Flanders P. The genetic properties of DNA transferred from ultraviolet-irradiated Hfr cells of Escherichia coli K-12 during mating. Genetics. 1968 Oct;60(2):243–255. doi: 10.1093/genetics/60.2.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Willetts N. S., Mount D. W. Genetic analysis of recombination-deficient mutants of Escherichia coli K-12 carrying rec mutations cotransducible with thyA. J Bacteriol. 1969 Nov;100(2):923–934. doi: 10.1128/jb.100.2.923-934.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Wood T. H. Effects of temperature, agitation, and donor strain on chromosome transfer in Escherichia coli K-12. J Bacteriol. 1968 Dec;96(6):2077–2084. doi: 10.1128/jb.96.6.2077-2084.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. YANOFSKY C., CARLTON B. C., GUEST J. R., HELINSKI D. R., HENNING U. ON THE COLINEARITY OF GENE STRUCTURE AND PROTEIN STRUCTURE. Proc Natl Acad Sci U S A. 1964 Feb;51:266–272. doi: 10.1073/pnas.51.2.266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Zipser D. Orientation of nonsense codons on the genetic map of the lac operon. Science. 1967 Sep 8;157(3793):1176–1177. doi: 10.1126/science.157.3793.1176. [DOI] [PubMed] [Google Scholar]
  64. de Haan P. G., Verhoef C. Genetic recombination in Escherichia coli. II. Calculation of incorporation frequency and relative map distance by recombinant analysis. Mutat Res. 1966 Apr;3(2):111–117. doi: 10.1016/0027-5107(66)90024-8. [DOI] [PubMed] [Google Scholar]

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

RESOURCES