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. 1979 Sep;139(3):859–865. doi: 10.1128/jb.139.3.859-865.1979

Genetic mapping of toxin regulatory mutations in Vibrio cholerae.

J J Mekalanos, R D Sublett, W R Romig
PMCID: PMC218032  PMID: 479110

Abstract

We have mapped a regulatory site mediating the hyperproduction of cholera toxin in mutants of Vibrio cholerae strain 569B. Mutations in this locus, called htx, result in the hypertoxinogenic phenotype, as measured by the ganglioside filter assay and immunoradial diffusion. Transposon-facilitated recombination was used to construct improved genetic donors in 569B parental and hypertoxinogenic mutant strains. Subsequent mapping by conjugation indicated that the htx locus was closely linked to the rif, str, and ilv loci of V. cholerae. Analysis of recombinants from these crosses suggested the following gene order: thy str htx rif ilv arg. The close genetic linkage of htx to rif (as high as 98%) resulted in a high comutation frequency of these two loci by nitrosoguanidine mutagenesis. Transfer of the htx mutant locus from a hypertoxinogenic donor to several unrelated Tox+ strains of V. cholerae caused a detectable elevation of toxin production in the recipients. These results suggest that toxin production in diverse strains of V. cholerae is controlled by a common regulatory mechanism in which the htx gene product plays a significant role.

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Selected References

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  1. Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baine W. B., Vasil M. L., Holmes R. K. Genetic mapping of mutations in independently isolated nontoxinogenic mutants of Vibrio cholerae. Infect Immun. 1978 Jul;21(1):194–200. doi: 10.1128/iai.21.1.194-200.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Callahan L. T., 3rd, Richardson S. H. Biochemistry of Vibrio cholerae virulence. 3. Nutritional requirements for toxin production and the effects of pH on toxin elaboration in chemically defined media. Infect Immun. 1973 Apr;7(4):567–572. doi: 10.1128/iai.7.4.567-572.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Finkelstein R. A., Vasil M. L., Holmes R. K. Studies on toxinogenesis in Vibrio cholerae. I. Isolation of mutants with altered toxinogenicity. J Infect Dis. 1974 Feb;129(2):117–123. doi: 10.1093/infdis/129.2.117. [DOI] [PubMed] [Google Scholar]
  5. Gerdes J. C., Romig W. R. Genetic basis of toxin production and pathogenesis in Vibrio cholerae: evidence against phage conversion. Infect Immun. 1975 Mar;11(3):445–452. doi: 10.1128/iai.11.3.445-452.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Guerola N., Ingraham J. L., Cerdá-Olmedo E. Induction of closely linked multiple mutations by nitrosoguanidine. Nat New Biol. 1971 Mar 24;230(12):122–125. doi: 10.1038/newbio230122a0. [DOI] [PubMed] [Google Scholar]
  7. Honda T., Finkelstein R. A. Selection and characteristics of a Vibrio cholerae mutant lacking the A (ADP-ribosylating) portion of the cholera enterotoxin. Proc Natl Acad Sci U S A. 1979 Apr;76(4):2052–2056. doi: 10.1073/pnas.76.4.2052. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Howard B. D. A prototype live oral cholera vaccine. Nature. 1971 Mar 12;230(5289):97–99. doi: 10.1038/230097a0. [DOI] [PubMed] [Google Scholar]
  9. Johnson S. R., Romig W. R. Transposon-facilitated recombination in Vibrio cholerae. Mol Gen Genet. 1979 Feb 16;170(1):93–101. doi: 10.1007/BF00268584. [DOI] [PubMed] [Google Scholar]
  10. Johnson S. R., Romig W. R. Vibrio cholerae hybrid sex factor that contains ampicillin transposon Tn1. J Bacteriol. 1979 Jan;137(1):531–536. doi: 10.1128/jb.137.1.531-536.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Levner M., Wiener F. P., Rubin B. A. Induction of Escherichia coli and Vibrio cholerae enterotoxins by an inhibitor of protein synthesis. Infect Immun. 1977 Jan;15(1):132–137. doi: 10.1128/iai.15.1.132-137.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mekalanos J. J., Collier R. J., Romig W. R. Affinity filters, a new approach to the isolation of tox mutants of Vibrio cholerae. Proc Natl Acad Sci U S A. 1978 Feb;75(2):941–945. doi: 10.1073/pnas.75.2.941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mekalanos J. J., Collier R. J., Romig W. R. Purification of cholera toxin and its subunits: new methods of preparation and the use of hypertoxinogenic mutants. Infect Immun. 1978 May;20(2):552–558. doi: 10.1128/iai.20.2.552-558.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mekalanos J. J., Collier R. J., Romig W. R. Simple method for purifying choleragenoid, the natural toxoid of Vibrio cholerae. Infect Immun. 1977 Jun;16(3):789–795. doi: 10.1128/iai.16.3.789-795.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pappenheimer A. M., Jr Diphtheria toxin. Annu Rev Biochem. 1977;46:69–94. doi: 10.1146/annurev.bi.46.070177.000441. [DOI] [PubMed] [Google Scholar]
  16. Parker C., Gauthier D., Tate A., Richardson K., Romig W. R. Expanded linkage map of Vibrio cholerae. Genetics. 1979 Feb;91(2):191–214. doi: 10.1093/genetics/91.2.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pierce N. F., Kaniecki E. A., Northrup R. S. Protection against experimental cholera by antitoxin. J Infect Dis. 1972 Dec;126(6):606–616. doi: 10.1093/infdis/126.6.606. [DOI] [PubMed] [Google Scholar]
  18. Richardson S. H. Factors influencing in vitro skin permeability factor production by Vibrio cholerae. J Bacteriol. 1969 Oct;100(1):27–34. doi: 10.1128/jb.100.1.27-34.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ruch F. E., Jr, Murphy J. R., Graf L. H., Field M. Isolation of nontoxinogenic mutants of Vibrio cholerae in a colorimetric assay for cholera toxin using the S49 mouse lymphosarcoma cell line. J Infect Dis. 1978 Jun;137(6):747–755. doi: 10.1093/infdis/137.6.747. [DOI] [PubMed] [Google Scholar]
  20. Vasil M. L., Holmes R. K., Finkelstein R. A. Conjugal transfer of a chromosomal gene determining production of enterotoxin in vibrio cholerae. Science. 1975 Mar 7;187(4179):849–850. doi: 10.1126/science.1114331. [DOI] [PubMed] [Google Scholar]

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