Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1983;2(10):1831–1838. doi: 10.1002/j.1460-2075.1983.tb01666.x

Construction of a correlated physical and genetic map of the Klebsiella pneumoniae hisDGO region using transposon Tn5 mutagenesis.

F J de Bruijn, I L Stroke, D J Marvel, F M Ausubel
PMCID: PMC555367  PMID: 6315405

Abstract

Multicopy plasmids containing the hisDG region of Klebsiella pneumoniae were mutagenized with transposon Tn5. The resulting plasmids were examined for their ability to complement hisD and hisG mutations in Escherichia coli. The physical location of Tn5 on each of the hisD::Tn5 and hisG::Tn5 plasmids was determined by restriction endonuclease analysis. By combining the two types of data, a precise correlated physical and genetic map of the K. pneumoniae hisDG region was constructed. Based on this analysis, the minimum sizes of the hisD and the hisG genes were calculated to be 1100 bp and 900 bp, respectively. The hisO(P) region was also identified. The insertional specificity of transposon Tn5 was shown to be very low. One unanticipated result was obtained: Tn5 insertions in the plasmid-borne hisG gene were not polar on hisD.

Full text

PDF
1831

Images in this article

1835Fig. 4.
on p.1835

Selected References

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

  1. Barnes W. M. DNA sequence from the histidine operon control region: seven histidine codons in a row. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4281–4285. doi: 10.1073/pnas.75.9.4281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berg D. E., Weiss A., Crossland L. Polarity of Tn5 insertion mutations in Escherichia coli. J Bacteriol. 1980 May;142(2):439–446. doi: 10.1128/jb.142.2.439-446.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bitar K. G., Firca J. R., Loper J. C. Histidinol dehydrogenase from salmonella typhimurium and Escherichia coli. Purification, some characteristics and the amino acid sequence around a reactive thiol group. Biochim Biophys Acta. 1977 Aug 23;493(2):429–440. doi: 10.1016/0005-2795(77)90199-4. [DOI] [PubMed] [Google Scholar]
  4. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  5. Bruni C. B., Musti A. M., Frunzio R., Blasi F. Structural and physiological studies of the Escherichia coli histidine operon inserted into plasmid vectors. J Bacteriol. 1980 Apr;142(1):32–42. doi: 10.1128/jb.142.1.32-42.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cannon F. C., Riedel G. E., Ausubel F. M. Overlapping sequences of Klebsiella pneumoniae nifDNA cloned and characterized. Mol Gen Genet. 1979 Jul 2;174(1):59–66. doi: 10.1007/BF00433306. [DOI] [PubMed] [Google Scholar]
  7. Cannon F. C., Riedel G. E., Ausubel F. M. Recombinant plasmid that carries part of the nitrogen fixation (nif) gene cluster of Klebsiella pneumoniae. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2963–2967. doi: 10.1073/pnas.74.7.2963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Di Nocera P. P., Blasi F., Di Lauro R., Frunzio R., Bruni C. B. Nucleotide sequence of the attenuator region of the histidine operon of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4276–4280. doi: 10.1073/pnas.75.9.4276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dixon R. A., Postgate J. R. Genetic transfer of nitrogen fixation from Klebsiella pneumoniae to Escherichia coli. Nature. 1972 May 12;237(5350):102–103. doi: 10.1038/237102a0. [DOI] [PubMed] [Google Scholar]
  14. Hakkaart M. J., Veltkamp E., Nijkamp H. J. Protein H encoded by plasmid Clo DF13 involved in lysis of the bacterial host. I. Localisation of the gene and identification and subcellular localisation of the gene H product. Mol Gen Genet. 1981;183(2):318–325. doi: 10.1007/BF00270635. [DOI] [PubMed] [Google Scholar]
  15. Hartman P. E., Hartman Z., Stahl R. C. Classification and mapping of spontaneous and induced mutations in the histidine operon of Salmonella. Adv Genet. 1971;16:1–34. doi: 10.1016/s0065-2660(08)60352-1. [DOI] [PubMed] [Google Scholar]
  16. Hayward G. S., Smith M. G. The chromosome of bacteriophage T5. I. Analysis of the single-stranded DNA fragments by agarose gel electrophoresis. J Mol Biol. 1972 Feb 14;63(3):383–395. doi: 10.1016/0022-2836(72)90435-4. [DOI] [PubMed] [Google Scholar]
  17. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  18. Johnson R. C., Yin J. C., Reznikoff W. S. Control of Tn5 transposition in Escherichia coli is mediated by protein from the right repeat. Cell. 1982 Oct;30(3):873–882. doi: 10.1016/0092-8674(82)90292-6. [DOI] [PubMed] [Google Scholar]
  19. Johnston H. M., Barnes W. M., Chumley F. G., Bossi L., Roth J. R. Model for regulation of the histidine operon of Salmonella. Proc Natl Acad Sci U S A. 1980 Jan;77(1):508–512. doi: 10.1073/pnas.77.1.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jorgensen R. A., Rothstein S. J., Reznikoff W. S. A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance. Mol Gen Genet. 1979;177(1):65–72. doi: 10.1007/BF00267254. [DOI] [PubMed] [Google Scholar]
  21. Kleckner N., Chan R. K., Tye B. K., Botstein D. Mutagenesis by insertion of a drug-resistance element carrying an inverted repetition. J Mol Biol. 1975 Oct 5;97(4):561–575. doi: 10.1016/s0022-2836(75)80059-3. [DOI] [PubMed] [Google Scholar]
  22. Kleckner N., Roth J., Botstein D. Genetic engineering in vivo using translocatable drug-resistance elements. New methods in bacterial genetics. J Mol Biol. 1977 Oct 15;116(1):125–159. doi: 10.1016/0022-2836(77)90123-1. [DOI] [PubMed] [Google Scholar]
  23. Kleckner N., Steele D. A., Reichardt K., Botstein D. Specificity of insertion by the translocatable tetracycline-resistance element Tn10. Genetics. 1979 Aug;92(4):1023–1040. doi: 10.1093/genetics/92.4.1023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kleckner N. Transposable elements in prokaryotes. Annu Rev Genet. 1981;15:341–404. doi: 10.1146/annurev.ge.15.120181.002013. [DOI] [PubMed] [Google Scholar]
  25. Klein R. D., Selsing E., Wells R. D. A rapid microscale technique for isolation of recombinant plasmid DNA suitable for restriction enzyme analysis. Plasmid. 1980 Jan;3(1):88–91. doi: 10.1016/s0147-619x(80)90037-2. [DOI] [PubMed] [Google Scholar]
  26. Kolter R., Yanofsky C. Attenuation in amino acid biosynthetic operons. Annu Rev Genet. 1982;16:113–134. doi: 10.1146/annurev.ge.16.120182.000553. [DOI] [PubMed] [Google Scholar]
  27. Laird A. J., Young I. G. Tn5 mutagenesis of the enterochelin gene cluster of Escherichia coli. Gene. 1980 Nov;11(3-4):359–366. doi: 10.1016/0378-1119(80)90075-x. [DOI] [PubMed] [Google Scholar]
  28. Lupski J. R., Smiley B. L., Blattner F. R., Godson G. N. Cloning and characterization of the Escherichia coli chromosomal region surrounding the dnaG Gene, with a correlated physical and genetic map of dnaG generated via transposon Tn5 mutagenesis. Mol Gen Genet. 1982;185(1):120–128. doi: 10.1007/BF00333800. [DOI] [PubMed] [Google Scholar]
  29. Merrick M., Filser M., Dixon R., Elmerich C., Sibold L., Houmard J. The use of translocatable genetic elements to construct a fine-structure map of the Klebsiella pneumoniae nitrogen fixation (nif) gene cluster. J Gen Microbiol. 1980 Apr;117(2):509–520. doi: 10.1099/00221287-117-2-509. [DOI] [PubMed] [Google Scholar]
  30. Miller J. H., Calos M. P., Galas D., Hofer M., Büchel D. E., Müller-Hill B. Genetic analysis of transpositions in the lac region of Escherichia coli. J Mol Biol. 1980 Nov 25;144(1):1–18. doi: 10.1016/0022-2836(80)90212-0. [DOI] [PubMed] [Google Scholar]
  31. Morgan E. A. Insertions of Tn 10 into an E. coli ribosomal RNA operon are incompletely polar. Cell. 1980 Aug;21(1):257–265. doi: 10.1016/0092-8674(80)90133-6. [DOI] [PubMed] [Google Scholar]
  32. Morrison D. A. Transformation and preservation of competent bacterial cells by freezing. Methods Enzymol. 1979;68:326–331. doi: 10.1016/0076-6879(79)68023-0. [DOI] [PubMed] [Google Scholar]
  33. Ow D. W., Ausubel F. M. Recombinant P4 bacteriophages propagate as viable lytic phages or as autonomous plasmids in Klebsiella pneumoniae. Mol Gen Genet. 1980;180(1):165–175. doi: 10.1007/BF00267366. [DOI] [PubMed] [Google Scholar]
  34. Pannekoek H., Hille J., Noordermeer I. Relief of polarity caused by transposon Tn5: application in mapping a cloned region of the Escherichia coli uvrB locus essential for UV resistance. Gene. 1980 Dec;12(1-2):51–61. doi: 10.1016/0378-1119(80)90015-3. [DOI] [PubMed] [Google Scholar]
  35. Piszkiewicz D., Tilley B. E., Rand-Meir T., Parsons S. M. Amino acid sequence of ATP phosphoribosyltransferase of Salmonella typhimurium. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1589–1592. doi: 10.1073/pnas.76.4.1589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Riedel G. E., Ausubel F. M., Cannon F. C. Physical map of chromosomal nitrogen fixation (nif) genes of Klebsiella pneumoniae. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2866–2870. doi: 10.1073/pnas.76.6.2866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Riedel G. E., Brown S. E., Ausubel F. M. Nitrogen fixation by Klebsiella pneumoniae is inhibited by certain multicopy hybrid nif plasmids. J Bacteriol. 1983 Jan;153(1):45–56. doi: 10.1128/jb.153.1.45-56.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rodriguez R. L., West R. W., Tait R. C., Jaynes J. M., Shanmugam K. T. Isolation and characterization of the hisG and hisD genes of Klebsiella pneumoniae. Gene. 1981 Dec;16(1-3):317–320. doi: 10.1016/0378-1119(81)90087-1. [DOI] [PubMed] [Google Scholar]
  39. Saedler H., Reif H. J., Hu S., Davidson N. IS2, a genetic element for turn-off and turn-on of gene activity in E. coli. Mol Gen Genet. 1974;132(4):265–289. doi: 10.1007/BF00268569. [DOI] [PubMed] [Google Scholar]
  40. Shanabruch W. G., Walker G. C. Localization of the plasmid (pKM101) gene(s) involved in recA+lexA+-dependent mutagenesis. Mol Gen Genet. 1980;179(2):289–297. doi: 10.1007/BF00425456. [DOI] [PubMed] [Google Scholar]
  41. Shaw K. J., Berg C. M. Escherichia coli K-12 auxotrophs induced by insertion of the transposable element Tn5. Genetics. 1979 Jul;92(3):741–747. doi: 10.1093/genetics/92.3.741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sommer H., Cullum J., Saedler H. IS2-43 and IS2-44: new alleles of the insertion sequence IS2 which have promoter activity. Mol Gen Genet. 1979 Aug;175(1):53–56. doi: 10.1007/BF00267855. [DOI] [PubMed] [Google Scholar]
  43. Stephens J. C., Artz S. W., Ames B. N. Guanosine 5'-diphosphate 3'-diphosphate (ppGpp): positive effector for histidine operon transcription and general signal for amino-acid deficiency. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4389–4393. doi: 10.1073/pnas.72.11.4389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Streicher S. L., Gurney E. G., Valentine R. C. The nitrogen fixation genes. Nature. 1972 Oct 27;239(5374):495–499. doi: 10.1038/239495a0. [DOI] [PubMed] [Google Scholar]
  45. Stüber D., Bujard H. Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci U S A. 1981 Jan;78(1):167–171. doi: 10.1073/pnas.78.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
  47. Verde P., Frunzio R., di Nocera P. P., Blasi F., Bruni C. B. Identification, nucleotide sequence and expression of the regulatory region of the histidine operon of Escherichia coli K-12. Nucleic Acids Res. 1981 May 11;9(9):2075–2086. doi: 10.1093/nar/9.9.2075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. West R. W., Jr, Rodriguez R. L. Construction and characterization of E. coli promoter-probe plasmid vectors. III. pBR322 derivatives with deletions in the tetracycline resistance promoter region. Gene. 1982 Dec;20(2):291–304. doi: 10.1016/0378-1119(82)90047-6. [DOI] [PubMed] [Google Scholar]
  49. de Bruijn F. J., Ausubel F. M. The cloning and transposon Tn5 mutagenesis of the glnA region of Klebsiella pneumoniae: identification of glnR, a gene involved in the regulation of the nif and hut operons. Mol Gen Genet. 1981;183(2):289–297. doi: 10.1007/BF00270631. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES