Abstract
The mini-Tn10 transposon (delta 16 delta 17Tn10) confers tetracycline resistance. When inserted between a gene and its promoter, it blocks transcription and prevents expression of that gene. Tetracycline in the medium induces divergent transcription of the tetA and tetR genes within the transposon, and this transcription extends beyond the transposon in both directions into the bacterial genes. If the mini-Tn10 inserts between an essential bacterial gene and its promoter, the insertion mutation can cause conditional growth which is dependent on the presence of tetracycline. Two essential genes in adjacent operons of Escherichia coli have been detected by screening for tetracycline dependence among tetracycline-resistant insertion mutants. These essential genes are the era gene in the rnc operon and the dpj gene in the adjacent pdxJ operon. The pdxJ operon has not been described previously. It consists of two genes, pdxJ and dpj. Whereas the dpj gene is essential for E. coli growth in all media tested, pdxJ is not essential. The pdxJ gene encodes a protein required in the biosynthesis of pyridoxine (vitamin B6).
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- Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bardwell J. C., Régnier P., Chen S. M., Nakamura Y., Grunberg-Manago M., Court D. L. Autoregulation of RNase III operon by mRNA processing. EMBO J. 1989 Nov;8(11):3401–3407. doi: 10.1002/j.1460-2075.1989.tb08504.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen S. M., Takiff H. E., Barber A. M., Dubois G. C., Bardwell J. C., Court D. L. Expression and characterization of RNase III and Era proteins. Products of the rnc operon of Escherichia coli. J Biol Chem. 1990 Feb 15;265(5):2888–2895. [PubMed] [Google Scholar]
- Chow W. Y., Berg D. E. Tn5tac1, a derivative of transposon Tn5 that generates conditional mutations. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6468–6472. doi: 10.1073/pnas.85.17.6468. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Crouch R. J. Ribonuclease 3 does not degrade deoxyribonucleic acid-ribonucleic acid hybrids. J Biol Chem. 1974 Feb 25;249(4):1314–1316. [PubMed] [Google Scholar]
- Dempsey W. B. Characterization of pyridoxine auxotrophs of Escherichia coli: P1 transduction. J Bacteriol. 1969 Mar;97(3):1403–1410. doi: 10.1128/jb.97.3.1403-1410.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dempsey W. B. Escherichia coli mutants tht require either pyridoxine or alanine. J Bacteriol. 1972 Sep;111(3):838–840. doi: 10.1128/jb.111.3.838-840.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Duester G. L., Holmes W. M. The distal end of the ribosomal RNA operon rrnD of Escherichia coli contains a tRNA1thr gene, two 5s rRNA genes and a transcription terminator. Nucleic Acids Res. 1980 Sep 11;8(17):3793–3807. doi: 10.1093/nar/8.17.3793. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hillen W., Schollmeier K., Gatz C. Control of expression of the Tn10-encoded tetracycline resistance operon. II. Interaction of RNA polymerase and TET repressor with the tet operon regulatory region. J Mol Biol. 1984 Jan 15;172(2):185–201. doi: 10.1016/s0022-2836(84)80037-6. [DOI] [PubMed] [Google Scholar]
- Inada T., Kawakami K., Chen S. M., Takiff H. E., Court D. L., Nakamura Y. Temperature-sensitive lethal mutant of era, a G protein in Escherichia coli. J Bacteriol. 1989 Sep;171(9):5017–5024. doi: 10.1128/jb.171.9.5017-5024.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kleckner N., Barker D. F., Ross D. G., Botstein D. Properties of the translocatable tetracycline-resistance element Tn10 in Escherichia coli and bacteriophage lambda. Genetics. 1978 Nov;90(3):427–461. doi: 10.1093/genetics/90.3.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kleckner N. DNA sequence analysis of Tn10 insertions: origin and role of 9 bp flanking repetitions during Tn10 translocation. Cell. 1979 Apr;16(4):711–720. doi: 10.1016/0092-8674(79)90087-4. [DOI] [PubMed] [Google Scholar]
- Lam H. M., Tancula E., Dempsey W. B., Winkler M. E. Suppression of insertions in the complex pdxJ operon of Escherichia coli K-12 by lon and other mutations. J Bacteriol. 1992 Mar;174(5):1554–1567. doi: 10.1128/jb.174.5.1554-1567.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lautenberger J. A., Court D., Papas T. S. High-level expression in Escherichia coli of the carboxy-terminal sequences of the avian myelocytomatosis virus (MC29) v-myc protein. Gene. 1983 Jul;23(1):75–84. doi: 10.1016/0378-1119(83)90218-4. [DOI] [PubMed] [Google Scholar]
- Lecocq J., Dambly C. A bacterial RNA polymerase mutant that renders lambda growth independent of the N and cro functions at 42 degrees C. Mol Gen Genet. 1976 Apr 23;145(1):53–64. doi: 10.1007/BF00331557. [DOI] [PubMed] [Google Scholar]
- March P. E., Lerner C. G., Ahnn J., Cui X., Inouye M. The Escherichia coli Ras-like protein (Era) has GTPase activity and is essential for cell growth. Oncogene. 1988 Jun;2(6):539–544. [PubMed] [Google Scholar]
- McKenney K., Shimatake H., Court D., Schmeissner U., Brady C., Rosenberg M. A system to study promoter and terminator signals recognized by Escherichia coli RNA polymerase. Gene Amplif Anal. 1981;2:383–415. [PubMed] [Google Scholar]
- Morgan W. D., Bear D. G., von Hippel P. H. Specificity of release by Escherichia coli transcription termination factor rho of nascent mRNA transcripts initiated at the lambda PR. J Biol Chem. 1984 Jul 10;259(13):8664–8671. [PubMed] [Google Scholar]
- Morrison P. T., Lovett S. T., Gilson L. E., Kolodner R. Molecular analysis of the Escherichia coli recO gene. J Bacteriol. 1989 Jul;171(7):3641–3649. doi: 10.1128/jb.171.7.3641-3649.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roa B. B., Connolly D. M., Winkler M. E. Overlap between pdxA and ksgA in the complex pdxA-ksgA-apaG-apaH operon of Escherichia coli K-12. J Bacteriol. 1989 Sep;171(9):4767–4777. doi: 10.1128/jb.171.9.4767-4777.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
- Schoenlein P. V., Roa B. B., Winkler M. E. Divergent transcription of pdxB and homology between the pdxB and serA gene products in Escherichia coli K-12. J Bacteriol. 1989 Nov;171(11):6084–6092. doi: 10.1128/jb.171.11.6084-6092.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simons R. W., Houman F., Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. doi: 10.1016/0378-1119(87)90095-3. [DOI] [PubMed] [Google Scholar]
- Takiff H. E., Chen S. M., Court D. L. Genetic analysis of the rnc operon of Escherichia coli. J Bacteriol. 1989 May;171(5):2581–2590. doi: 10.1128/jb.171.5.2581-2590.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor D. P., Greenberg J., Rownd R. H. Generation of miniplasmids from copy number mutants of the R plasmid NR1. J Bacteriol. 1977 Dec;132(3):986–995. doi: 10.1128/jb.132.3.986-995.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Way J. C., Davis M. A., Morisato D., Roberts D. E., Kleckner N. New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene. 1984 Dec;32(3):369–379. doi: 10.1016/0378-1119(84)90012-x. [DOI] [PubMed] [Google Scholar]
- d'Aubenton Carafa Y., Brody E., Thermes C. Prediction of rho-independent Escherichia coli transcription terminators. A statistical analysis of their RNA stem-loop structures. J Mol Biol. 1990 Dec 20;216(4):835–858. doi: 10.1016/s0022-2836(99)80005-9. [DOI] [PubMed] [Google Scholar]