1887

Abstract

SUMMARY: The DNA primase gene of the promiscuous IncP-1 conjugative plasmid RP1, encoding two polypeptides of 118 and 80 kDa, was inserted into the transposon Tn5 in The derivative transposon, Tn2523, was then transposed to a temperature-sensitive replication mutant of the promiscuous IncP-1 conjugative plasmid R68 at permissive temperature and the plasmid transferred to strain PAO. The latter strain was then grown at non-permissive temperature to identify transposition of Tn2523 into the chromosome. Immunological and enzymic analysis showed the expression of functional primase polypeptides in the constructed strain. This strain also restored wild-type conjugational transfer proficiency, by complementation, to mutants of the IncP-1 plasmid R18 affected in transfer from to or to due to transposon Tn7 insertion mutations in the primase gene. This strategy of cloning into a transposon and integration into the bacterial chromosome should facilitate genetic manipulation and studies of gene expression in a range of Gram-negative bacteria.

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1986-03-01
2021-10-20
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References

  1. Bachman K., Ptashne M., Gilbert W. 1976; Construction of plasmids carrying the Cl gene of bacteriophage λ. Proceedings of the National Academy of Sciences of the United States of America 73:4174–4178
    [Google Scholar]
  2. Bachmann B. J. 1983; Linkage map of Escherichia coli. Edition 7. Microbiological Reviews 47:180–230
    [Google Scholar]
  3. Bagdasarian M. M., Amann E., Lurz R., Ruckert B., Bagdasarian M. 1983; Activity of the hybrid trp-lac (tac) promoter of Escherichia coli in Pseudomonas putida: construction of broad-host-range, controlled-expression vectors. Gene 26:273–282
    [Google Scholar]
  4. Bolivar M. M., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Croser J., Falkow S. 1977; Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113
    [Google Scholar]
  5. Boulnois G. J., Wilkins B. M. 1979; novel priming system for conjugational synthesis of an Incl𝛼 plasmid in recipients. Molecular and General Genetics 175:275–279
    [Google Scholar]
  6. Burkhardt H., Riess G., Puhler A. 1979; Relationship of group PI plasmids revealed by heteroduplex experiments: RP1, RP4, R68 and RK2 are identical. Journal of General Microbiology 114:341–348
    [Google Scholar]
  7. Chang A. C. Y., Cohen S. N. 1978; Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic plasmid. Journal of General Bacteriology 134:1141–1156
    [Google Scholar]
  8. Cohen S. N., Chang A. C. Y., Hsu L. 1972; Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proceedings of the National Academy of Sciences of the United States of America 69:2110–2114
    [Google Scholar]
  9. Cowan P., Krishnapillai V. 1982; Tn7 insertion mutations affecting the host range of the promiscuous IncP-1 plasmid R68. Plasmid 8:164–174
    [Google Scholar]
  10. Finger J., Krishnapillai V. 1980; Host range, entry exclusion and incompatibility of Pseudomonas aeruginosa FP plasmids. Plasmid 3:332–342
    [Google Scholar]
  11. Frey J., Bagdasarian M., Feiss D., Franklin. F. C. H., Deshusses J. 1983; Stable cosmid vectors that enable the introduction of cloned fragments into a wide range of Gram-negative bacteria. Gene 24:299–308
    [Google Scholar]
  12. Friedman A. M., Long S. R., Brown S. E., Buikma W. J., Ausubel F. M. 1982; Construction of a broad host range cosmid cloning vector and its use in the genetic analvsis of Rhizobium mutants. Gene 18:289–296
    [Google Scholar]
  13. Grinsted J., Bennett P. M., Higginson S., Richmond M. H. 1978; Regional preference of insertion of Tn501 and Tn802 into RP1 and its derivatives. Molecular and General Genetics 166:313–320
    [Google Scholar]
  14. Grinter N. J. 1983; A broad-host-range cloning vector transposable to various replicons. Gene 21:133–143
    [Google Scholar]
  15. Haas D., Holloway B. W., Schambock A., Leisinger T. 1977; The genetic organization of arginine biosynthesis in Pseudomonas aeruginosa. Molecular and General Genetics 154:7–22
    [Google Scholar]
  16. Haas D., CryZ S. J., Jr, Itoh Y., Leisinger T., Luthi E., Mercenier A., Reimann C., Rella M., Soldati L., Watson J. M., Wretlind B. 1984; Some applications of transposon insertion mutagenesis in Pseudomonas. Edited by H. Heslot. Paris: Societe Francaise de Microbiologie. In Genetique des Microorganismes Industrieles pp:91–111
    [Google Scholar]
  17. Holloway B. W. 1965; Variations in restriction and modification following increase of growth temperature of Pseudomonas aeruginosa. Virology 25:634–642
    [Google Scholar]
  18. Holloway B. W., Matsumoto M. 1984; Pseudomonas aeruginosa. Edited by S. J. O’Brien. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory. In Genetic Maps 1984 pp:194–197
    [Google Scholar]
  19. Holloway B. W., Crowther C., Dean H., Hage-dorn J., Holmes N., Morgan A. F. 1982; Integration of plasmids into the Pseudomonas chromosome. Edited by S. Mitsuhashi. Tokyo: Japan Scientific Societies Press. In Drug Resistance in Bacteria, pp:231–242
    [Google Scholar]
  20. Isaac J. H., Holloway B. W. 1968; Control of pyrimidine biosynthesis in Pseudomonas aeruginosa. Journal of Bacteriology 96:1732–1741
    [Google Scholar]
  21. Kim K., Meyer R. J. 1984; Novel system for recognizing and eliminating foreign DNA in Pseudomonas putida. Journal of Bacteriology 159:678–682
    [Google Scholar]
  22. Krishnapillai V., Royle P., Lehrer J. 1981; Insertions of the transposon Tnl into the Pseudomonas aeruginosa chromosome. Genetics 97:495–511
    [Google Scholar]
  23. Krishnapillai V., Nash J., Lanka E. 1984; Insertion mutations in the promiscuous Inc P-1 plasmid R18 which affect its host range between Pseudomonas species. Plasmid 12:170–180
    [Google Scholar]
  24. Lanka E., Barth P. T. 1981; Plasmid RP4 specifies a deoxyribonucleic acid primase involved in its conjugal transfer and maintenance. Journal of Bacteriology 148:769–781
    [Google Scholar]
  25. Lanka E., Lurz R., Furste J. P. 1983; Molecular cloning and mapping of SphI restriction fragments of plasmid RP4. Plasmid 10:303–307
    [Google Scholar]
  26. Lanka E., Lurz R., Kroger M., Furste J. P. 1984; Plasmid RP4 encodes two forms of a DNA primase. Molecular and General Genetics 194:65–72
    [Google Scholar]
  27. Mandel M., Higa A. 1970; Calcium dependent bacteriophage DNA infection. Journal of Molecular Biology 53:159–162
    [Google Scholar]
  28. Maniatis T., Fritsch E. F., Sambrook J. 1982; Cold Spring Harbor, NY: Cold Spring Harbor Laboratory. Molecular Cloning: A Laboratory Manual.
    [Google Scholar]
  29. O'Hoy K., Krishnapillai V. 1982; Transposon mutagenesis of the Pseudomonas aeruginosa PAG chromosome and the isolation of high frequency recombination donors. FEMS Microbiology Letters 29:299–303
    [Google Scholar]
  30. Prozorov A. A., Bashkirov V. I., Khasanov F. K., Glumova E. F., Irich V. Y. 1985; Insertion of eukaryotic DNA into the Bacillus subtilis genome by means of a temperature-sensitive plasmid vector. Gene 34:39–46
    [Google Scholar]
  31. Rothstein S. J., Reznikoff W. S. 1981; The functional differences in the inverted repeats of Tn5 are caused by a single base pair non-homology. Cell 23:191–199
    [Google Scholar]
  32. Schilf W., Krishnapillai V. 1985; Genetic basis of non-transposition of Tn5 in Pseudomonas aeruginosa following mobilization of RP4Mob: :Tn5 from Escherichia coli. FEMS Microbiology Letters 28:351–354
    [Google Scholar]
  33. Simon R., Priefer U., Puhler A. 1983a; Vector plasmids for in vivo and in vitro manipulations of Gram-negative bacteria. Edited by A. Puhler. Berlin: Springer-Verlag. In Molecular Genetics of the Bacteria-Plant Interaction, pp:98–106
    [Google Scholar]
  34. Simon R., Priefer U., Puhler A. 1983b; A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gramnegative bacteria. Bio/Technology 1:784–791
    [Google Scholar]
  35. Smith C. A., Thomas C. M. 1985; Comparison of the nucleotide sequences of the vegetative replication origins of broad host range IncP plasmids R751 and RK2 reveals conserved features of probable functional importance. Nucleic Acids Research 13:557–572
    [Google Scholar]
  36. Stokes H. W., Moore R. J., Krishnapillai V. 1985; Complementation analysis in Pseudomonas aeruginosa of the transfer genes of the wide host range R plasmid R18. Plasmid 5:202–212
    [Google Scholar]
  37. Towner K. J. 1978; Chromosome mapping in Acinetobacter calcoaceticus. Journal of General Microbiology 104:175–180
    [Google Scholar]
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