Summary
Five gene products required for Tn7 transposition were identified using genetic complementation tests. Four of these (tns A, tnsB, tnsC and tnsD) are essential for insertion into the attachment site, whereas the fifth (tnsE) is required for transposition to plasmids which lack this site. tnsD is not required for transposition to plasmids lacking the attachment site. This analysis used a chloramphenicol-resistant ‘mini’ transposon containing Tn7 termini but no complete Tn7 gene product and several compatible expression vectors containing cloned Tn7 fragments. The use of transcriptional and translational fusions allowed the identification of two promoters (P1 and P2) at the right-hand end of the transposon and indicated that at least tnsA, tnsB and tnsC are translated in the same direction. Expression from P1 appears to be repressed by tnsB.
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References
Arthur A, Sherratt D (1979) Dissection of the transposition process: A transposon-encoded site-specific recombination system. Mol Gen Genet 175:267–274
Barth PT, Datta N, Hedges RW, Grinter NJ (1976) Transposition of a deoxyribonucleic acid sequence encoding trimethoprim and streptomycin resistance from R483 to other replicons. J Bacteriol 125:800–810
Barth PT, Grinter NJ (1977) Map of RP4 derived by insertion of Transposon C. J Mol Biol 113:455–474
Barth PT, Grinter NJ, Bradley DE (1978) Conjugal transfer system of plasmid RP4: Analysis by Transposon 7 insertion. J Bacteriol 133:43–52
Birmboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
Boyd AC, Sherratt DJ (1986) Polar mobilisation of the Escherichia coli chromosome by the ColE1 transfer origin. Mol Gen Genet 203:496–504
Brevet J, Faure, F, Borowski D (1985) Tn7-encoded proteins. Mol Gen Genet 201:258–264
Caruso M, Shapiro J (1982) Interaction of Tn7 and temperate phage F116L of Pseudomonas aeruginosa. Mol Gen Genet 188:292–298
Chang ACY, Cohen SN (1978) Construction and characterisation of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic mini plasmid. J Bacteriol 134:1141–1156
Datta N, Hedges RW (1972) Trimethoprim resistance conferred by W plasmids on Enterobacteriaceae. J Gen Microbiol 72:349–355
Ely B (1982) Transposition of Tn7 occurs at a single site on the Caulobacter crescentus chromosome. J Bacteriol 151:1056–1058
Gay N, Tybulewicz VLJ, Walker JE (1986) Insertion of transposon Tn7 into E. coli. Biochem J 234:111–117
Gosti-Testu F, Norris V, Brevet J (1983) Restriction map of Tn7. Plasmid 10:96–99
Hauer B, Shapiro J (1983) Control of Tn7 Transposition. Mol Gen Genet 194:149–158
Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197
Horii Z, Clark AS (1973) Genetic analysis of the recF pathway to genetic recombination in Escherichia coli K12: Isolation and characterisation of mutants. J Mol Biol 80: 327–344
Kennedy CK (1971) Induction of colicin production by high temperature or inhibition of protein synthesis. J Bacteriol 108:10–19
Kleckner N (1981) Transposable elements in prokaryotes. Annu Rev Genet 15:341–404
Lichtenstein C, Brenner S (1981) Site-specific properties of Tn7 transposition into the E. coli chromosome. Mol Gen Genet 183:380–387
Lichtenstein C, Brenner S (1982) Unique insertion site of Tn7 in the E. coli chromosome. Nature 297:601–603
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, New York
McKenney K, Shimatake H, Court D, Schmeissner U, Brady C, Rosenberg M (1981) A system to study promoter and terminator signals recognised by Escherichia coli RNA polymerase. In: Chirikjian JG, Papas TS (eds) Gene amplification and analysis. vol II. Elsevier, North Holland, Amsterdam, pp 385–415
Messing J, Gronenborn B, Muller-Hill B, Hofschneider PH (1977) Filamentous coliphage M13 as a cloning vehicle: Insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro. Proc Natl Acad Sci USA 74:3642–3646
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, New York
Minton NP, (1984) Improved plasmid vectors for the isolation of translational lac gene fusions. Gene 31:269–273
Moore RJ, Krishnapillai V (1982) Tn7 and TN501 Insertions into Pseudomonas aeruginosa plasmid R91-5: Mapping of two transfer regions. J Bacteriol 149:276–283
Nomark S, Bergstrom S, Edlund T, Grundstrom T, Jaurin B, Frederik PL, Olsson O (1983) Overlapping genes. Annu Rev Genet 17:499–525
Oka A, Sugisaki H, Takanami M (1981) Nucleotide sequence of the kanamycin resistance transposon Tn903. J Mol Biol 147:217–226
Ouartsi A, Borowski D, Brevet J (1985) Genetic analysis of Tn7 transposition. Mol Gen Genet 198:221–227
Smith GM, Jones P (1984) Effects of deletions in the transposon Tn7 on its frequency of transposition. J Bacteriol 157:962–964
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Thomson JA, Hendson M, Magnes RM (1981) Mutagenesis by insertion of drug resistance transposon into a Vibrio species. J Bacteriol 148:374–378
Turner P, Barber C, Daniels M (1984) Behaviour of the transposons Tn5 and Tn7 in Xanthomonas campestris pv campestris. Mol Gen Genet 195:101–107
Vieira J, Messing J (1982) The pUC plasmid, an M13mp7 derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:529–268
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Rogers, M., Ekaterinaki, N., Nimmo, E. et al. Analysis of Tn7 transposition. Mol Gen Genet 205, 550–556 (1986). https://doi.org/10.1007/BF00338097
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DOI: https://doi.org/10.1007/BF00338097