Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1991 Aug;173(15):4675–4682. doi: 10.1128/jb.173.15.4675-4682.1991

Molecular cloning of two linked loci that increase the transformability of transformation-deficient mutants of Haemophilus influenzae.

T G Larson 1, E Roszczyk 1, S H Goodgal 1
PMCID: PMC208144  PMID: 1649818

Abstract

A plasmid containing a 13.3-kb insert (pER194) was isolated from an EcoRI genomic library of Haemophilus influenzae on the basis of its ability to increase the transformability of the transformation-deficient mutants Com-78 and Com-101. The plasmid failed to increase the transformability of the Rec-1 and Rec-2 mutants, indicating that the mutations producing the Com-78 and Com-101 phenotypes are distinct from those giving rise to the Rec-1 and Rec-2 phenotypes. The physical mapping of the cloned fragment on the H. influenzae chromosome was found to be consistent with the genetic mapping of the Com-101 trait. A 2.8-kb EcoRI-BglII subfragment, representing one end of the 13.3-kb clone, was found to increase the transformation frequency of the Com-78 and Com-101 mutants when supplied in trans, indicating that the subfragment carries one or more loci required for chromosomal transformation. The corresponding region of the Com-101 chromosome was determined by hybridization analysis to contain a 0.3-kb insertion, suggesting that the Com-101 strain may contain an insertion mutation at this locus. A 3.0-kb EcoRI-MluI subfragment, representing the other end of the 13.3-kb EcoRI fragment, was found to increase the transformation frequency of the Com-101 mutant but not of the Com-78 mutant, suggesting that the Com-101 phenotype results from a complex genotype involving mutations at two or more transformation-related loci. This conclusion is consistent with data indicating that the Com-101 trait can be genetically separated into at least two components.

Full text

PDF
4675

Images in this article

4679Image
on p.4679

Selected References

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

  1. ALEXANDER H. E., LEIDY G. Determination of inherited traits of H. influenzae by desoxyribonucleic acid fractions isolated from type-specific cells. J Exp Med. 1951 Apr 1;93(4):345–359. doi: 10.1084/jem.93.4.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albano M., Breitling R., Dubnau D. A. Nucleotide sequence and genetic organization of the Bacillus subtilis comG operon. J Bacteriol. 1989 Oct;171(10):5386–5404. doi: 10.1128/jb.171.10.5386-5404.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BARNHART B. J., HERRIOTT R. M. PENETRATION OF DEOXYRIBONUCLEIC ACID INTO HEMOPHILUS INFLUENZAE. Biochim Biophys Acta. 1963 Sep 17;76:25–39. [PubMed] [Google Scholar]
  4. BERNS K. I., THOMAS C. A., Jr ISOLATION OF HIGH MOLECULAR WEIGHT DNA FROM HEMOPHILUS INFLUENZAE. J Mol Biol. 1965 Mar;11:476–490. doi: 10.1016/s0022-2836(65)80004-3. [DOI] [PubMed] [Google Scholar]
  5. Balganesh M., Arrigoni L., Setlow J. K. Plasmid-to-chromosome gene transfer in Haemophilus influenza during growth. J Bacteriol. 1986 Oct;168(1):458–459. doi: 10.1128/jb.168.1.458-459.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Balganesh M., Setlow J. K. Differential behavior of plasmids containing chromosomal DNA insertions of various sizes during transformation and conjugation in Haemophilus influenzae. J Bacteriol. 1985 Jan;161(1):141–146. doi: 10.1128/jb.161.1.141-146.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Barcak G. J., Tomb J. F., Laufer C. S., Smith H. O. Two Haemophilus influenzae Rd genes that complement the recA-like mutation rec-1. J Bacteriol. 1989 May;171(5):2451–2457. doi: 10.1128/jb.171.5.2451-2457.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Barouki R., Smith H. O. Reexamination of phenotypic defects in rec-1 and rec-2 mutants of Haemophilus influenzae Rd. J Bacteriol. 1985 Aug;163(2):629–634. doi: 10.1128/jb.163.2.629-634.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Beattie K. L., Setlow J. K. Transformation-defective strains of Haemophilus influenzae. Nat New Biol. 1971 Jun 9;231(23):177–179. doi: 10.1038/newbio231177a0. [DOI] [PubMed] [Google Scholar]
  10. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Caster J. H., Postel E. H., Goodgal S. H. Competence mutants: isolation of transformation deficient strains of Haemophilus influenzae. Nature. 1970 Aug 1;227(5257):515–517. doi: 10.1038/227515a0. [DOI] [PubMed] [Google Scholar]
  12. Chandler M. S., Morrison D. A. Competence for genetic transformation in Streptococcus pneumoniae: molecular cloning of com, a competence control locus. J Bacteriol. 1987 May;169(5):2005–2011. doi: 10.1128/jb.169.5.2005-2011.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. GOODGAL S. H., HERRIOTT R. M. Studies on transformations of Hemophilus influenzae. I. Competence. J Gen Physiol. 1961 Jul;44:1201–1227. doi: 10.1085/jgp.44.6.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Herriott R. M., Meyer E. M., Vogt M. Defined nongrowth media for stage II development of competence in Haemophilus influenzae. J Bacteriol. 1970 Feb;101(2):517–524. doi: 10.1128/jb.101.2.517-524.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Joshi V. P., Notani N. K. Cloning of novr gene in Haemophilus influenzae. Indian J Exp Biol. 1984 Dec;22(12):625–628. [PubMed] [Google Scholar]
  16. Kauc L., Goodgal S. H. Introduction of transposon Tn916 DNA into Haemophilus influenzae and Haemophilus parainfluenzae. J Bacteriol. 1989 Dec;171(12):6625–6628. doi: 10.1128/jb.171.12.6625-6628.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kauc L., Mitchell M., Goodgal S. H. Size and physical map of the chromosome of Haemophilus influenzae. J Bacteriol. 1989 May;171(5):2474–2479. doi: 10.1128/jb.171.5.2474-2479.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kooistra J., Venema G. Fate of donor DNA in some poorly transformable strains of Haemophilus influenzae. Mutat Res. 1970 Mar;9(3):245–253. doi: 10.1016/0027-5107(70)90126-0. [DOI] [PubMed] [Google Scholar]
  19. Kooistra J., Venema G. Fate of donor deoxyribonucleic acid in a highly transformation-deficient strain of Haemophilus influenzae. J Bacteriol. 1974 Sep;119(3):705–717. doi: 10.1128/jb.119.3.705-717.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Larson T. G., Goodgal S. H. Sequence and transcriptional regulation of com101A, a locus required for genetic transformation in Haemophilus influenzae. J Bacteriol. 1991 Aug;173(15):4683–4691. doi: 10.1128/jb.173.15.4683-4691.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lee J. J., Smith H. O., Redfield R. J. Organization of the Haemophilus influenzae Rd genome. J Bacteriol. 1989 Jun;171(6):3016–3024. doi: 10.1128/jb.171.6.3016-3024.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McCarthy D. Cloning of the rec-2 locus of Haemophilus influenzae. Gene. 1989 Jan 30;75(1):135–143. doi: 10.1016/0378-1119(89)90390-9. [DOI] [PubMed] [Google Scholar]
  23. Michalka J., Goodgal S. H. Genetic and physical map of the chromosome of Hemophilus influenzae. J Mol Biol. 1969 Oct 28;45(2):407–421. doi: 10.1016/0022-2836(69)90115-6. [DOI] [PubMed] [Google Scholar]
  24. Notani N. K., Setlow J. K., Joshi V. R., Allison D. P. Molecular basis for the transformation defects in mutants of Haemophilus influenzae. J Bacteriol. 1972 Jun;110(3):1171–1180. doi: 10.1128/jb.110.3.1171-1180.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Postel E. H., Goodgal S. H. Competence mutants. 3. Responses to radiations. J Bacteriol. 1972 Jan;109(1):298–306. doi: 10.1128/jb.109.1.298-306.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Postel E. H., Goodgal S. H. Uptake of "single-stranded" DNA in Hemophilus influenzae and its ability to transform. J Mol Biol. 1966 Apr;16(2):317–327. doi: 10.1016/s0022-2836(66)80175-4. [DOI] [PubMed] [Google Scholar]
  27. Setlow J. K., Boling M. E., Beattie K. L., Kimball R. F. A complex of recombination and repair genes in Haemophilus influenzae. J Mol Biol. 1972 Jul 21;68(2):361–378. doi: 10.1016/0022-2836(72)90218-5. [DOI] [PubMed] [Google Scholar]
  28. Setlow J. K., Brown D. C., Boling M. E., Mattingly A., Gordon M. P. Repair of deoxyribonucleic acid in Haemophilus influenzae. I. X-ray sensitivity of ultraviolet-sensitive mutants and their behavior as hosts to ultraviolet-irradiated bacteriophage and transforming deoxyribonucleic acid. J Bacteriol. 1968 Feb;95(2):546–558. doi: 10.1128/jb.95.2.546-558.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Setlow J. K., Spikes D., Griffin K. Characterization of the rec-1 gene of Haemophilus influenzae and behavior of the gene in Escherichia coli. J Bacteriol. 1988 Sep;170(9):3876–3881. doi: 10.1128/jb.170.9.3876-3881.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Stuy J. H. Cloning and characterization of the Haemophilus influenzae Rd rec-1+ gene. J Bacteriol. 1989 Aug;171(8):4395–4401. doi: 10.1128/jb.171.8.4395-4401.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Stuy J. H. Transfer of genetic information within a colony of Haemophilus influenzae. J Bacteriol. 1985 Apr;162(1):1–4. doi: 10.1128/jb.162.1.1-4.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Stuy J. H., Walter R. B. Effect of glycerol on plasmid transfer in genetically competent Haemophilus influenzae. Mol Gen Genet. 1986 May;203(2):296–299. doi: 10.1007/BF00333969. [DOI] [PubMed] [Google Scholar]
  33. Tomb J. F., Barcak G. J., Chandler M. S., Redfield R. J., Smith H. O. Transposon mutagenesis, characterization, and cloning of transformation genes of Haemophilus influenzae Rd. J Bacteriol. 1989 Jul;171(7):3796–3802. doi: 10.1128/jb.171.7.3796-3802.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Weinrauch Y., Guillen N., Dubnau D. A. Sequence and transcription mapping of Bacillus subtilis competence genes comB and comA, one of which is related to a family of bacterial regulatory determinants. J Bacteriol. 1989 Oct;171(10):5362–5375. doi: 10.1128/jb.171.10.5362-5375.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES