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. 1981 Jul;33(1):142–148. doi: 10.1128/iai.33.1.142-148.1981

Genetic mapping of chromosomal determinants for the production of the exopolysaccharide alginate in a Pseudomonas aeruginosa cystic fibrosis isolate.

D E Ohman, A M Chakrabarty
PMCID: PMC350668  PMID: 6790439

Abstract

Mucoid Pseudomonas aeruginosa strain FRD, a sputum isolate from a cystic fibrosis patient, was used to develop a genetic system. The mucoid appearance is due to the biosynthesis of the exopolysaccharide alginate and is a potential virulence factor of the organism. The sex factor plasmid FP2 was used for uninterrupted genetic exchange to investigate the nature of spontaneous mutations which produce frequent alginate-negative (Alg-) derivatives. Crosses between Alg+ donors and Alg- recipients demonstrated linkage between alginate genes and chromosomal markers. Crosses between an Alg- donor and Alg- recipients produced Alg+ recombinants at frequencies that varied, depending on the recipient strains used. This indicated that more than one genetic locus was associated with spontaneous mutation leading to loss of the mucoid character. Three classes of Alg- mutants were identified. Genetic exchange experiments showed that the loci of the alginate (alg) mutations of the three mutant classes are in the same region of the chromosome. The sex factor plasmid R68.45 was used for nonpolarized chromosome transfer and demonstrated close linkage between chromosomal markers (his-1, met-1) and alg markers. This was consistent with the data obtained in FP2-mediated crosses. Thus, the evidence obtained indicated that the alg genes which undergo frequent mutation are chromosomal, that several loci are involved, and that these alg loci are apparently clustered on the chromosome.

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Selected References

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

  1. Diaz E., Mosovich L. L., Neter E. Serogroups of Pseudomonas aeruginosa and the immune response of patients with cystic fibrosis. J Infect Dis. 1970 Mar;121(3):269–274. doi: 10.1093/infdis/121.3.269. [DOI] [PubMed] [Google Scholar]
  2. Doggett R. G. Incidence of mucoid Pseudomonas aeruginosa from clinical sources. Appl Microbiol. 1969 Nov;18(5):936–937. doi: 10.1128/am.18.5.936-937.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Evans L. R., Linker A. Production and characterization of the slime polysaccharide of Pseudomonas aeruginosa. J Bacteriol. 1973 Nov;116(2):915–924. doi: 10.1128/jb.116.2.915-924.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fyfe J. A., Govan J. R. Alginate synthesis in mucoid Pseudomonas aeruginosa: a chromosomal locus involved in control. J Gen Microbiol. 1980 Aug;119(2):443–450. doi: 10.1099/00221287-119-2-443. [DOI] [PubMed] [Google Scholar]
  5. Govan J. R., Fyfe J. A., McMillan C. The instability of mucoid Pseudomonas aeruginosa: fluctuation test and improved stability of the mucoid form in shaken culture. J Gen Microbiol. 1979 Jan;110(1):229–232. doi: 10.1099/00221287-110-1-229. [DOI] [PubMed] [Google Scholar]
  6. Govan J. R. Mucoid strains of Pseudomonas aeruginosa: the influence of culture medium on the stability of mucus production. J Med Microbiol. 1975 Nov;8(4):513–522. doi: 10.1099/00222615-8-4-513. [DOI] [PubMed] [Google Scholar]
  7. Haas D., Holloway B. W. Chromosome mobilization by the R plasmid R68.45: a tool in Pseudomonas genetics. Mol Gen Genet. 1978 Jan 17;158(3):229–237. doi: 10.1007/BF00267194. [DOI] [PubMed] [Google Scholar]
  8. Haas D., Holloway B. W., Schamböck A., Leisinger T. The genetic organization of arginine biosynthesis in Pseudomonas aeruginosa. Mol Gen Genet. 1977 Jul 7;154(1):7–22. doi: 10.1007/BF00265571. [DOI] [PubMed] [Google Scholar]
  9. Holloway B. W., Krishnapillai V., Morgan A. F. Chromosomal genetics of Pseudomonas. Microbiol Rev. 1979 Mar;43(1):73–102. doi: 10.1128/mr.43.1.73-102.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Knutson C. A., Jeanes A. A new modification of the carbazole analysis: application to heteropolysaccharides. Anal Biochem. 1968 Sep;24(3):470–481. doi: 10.1016/0003-2697(68)90154-1. [DOI] [PubMed] [Google Scholar]
  11. Linker A., Jones R. S. A new polysaccharide resembling alginic acid isolated from pseudomonads. J Biol Chem. 1966 Aug 25;241(16):3845–3851. [PubMed] [Google Scholar]
  12. Markowitz S. M., Macrina F. L., Phibbs P. V., Jr R-factor inheritance and plasmid content in mucoid Pseudomonas aeruginosa. Infect Immun. 1978 Nov;22(2):530–539. doi: 10.1128/iai.22.2.530-539.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Marks M. I., Prentice R., Swarson R., Cotton E. K., Eickhoff T. C. Carbenicillin and gentamicin: pharmacologic studies in patients with cystic fibrosis and pseudomonas pulmonary infections. J Pediatr. 1971 Nov;79(5):822–828. doi: 10.1016/s0022-3476(71)80401-8. [DOI] [PubMed] [Google Scholar]
  14. Martin D. R. Mucoid variation in Pseudomonas aeruginosa induced by the action of phage. J Med Microbiol. 1973 Feb;6(1):111–118. doi: 10.1099/00222615-6-1-111. [DOI] [PubMed] [Google Scholar]
  15. Mian F. A., Jarman T. R., Righelato R. C. Biosynthesis of exopolysaccharide by Pseudomonas aeruginosa. J Bacteriol. 1978 May;134(2):418–422. doi: 10.1128/jb.134.2.418-422.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Reynolds H. Y., Di Sant'Agnese P. A., Zierdt C. H. Mucoid Pseudomonas aeruginosa. A sign of cystic fibrosis in young adults with chronic pulmonary disease? JAMA. 1976 Nov 8;236(19):2190–2192. doi: 10.1001/jama.236.19.2190. [DOI] [PubMed] [Google Scholar]
  17. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  18. Watson J. M., Holloway B. W. Chromosome mapping in Pseudomonas aeruginosa PAT. J Bacteriol. 1978 Mar;133(3):1113–1125. doi: 10.1128/jb.133.3.1113-1125.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Zierdt C. H., Schmidt P. J. Dissociation in Pseudomonas aeruginosa. J Bacteriol. 1964 May;87(5):1003–1010. doi: 10.1128/jb.87.5.1003-1010.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. di Sant'Agnese P. A., Davis P. B. Research in cystic fibrosis (third of three parts). N Engl J Med. 1976 Sep 9;295(11):597–602. doi: 10.1056/NEJM197609092951105. [DOI] [PubMed] [Google Scholar]

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