Summary
The conjugative shuttle transposon Tn1545 from Streptococcus pneumoniae transposes in various gram-positive bacterial genera following self-transfer and in Escherichia coli after cloning. Analysis of the junction fragments and of the targets before insertion and after excision of the element by DNA hybridization and sequencing indicated that Tn1545 (1) is not flanked by terminal repeated sequences in either direct or opposite orientation, (2) is flanked, in an asymmetric fashion, by terminal variable base pairs, one at the left and three at the right of the element, (3) inserts in a target DNA consensus sequence, (4) does not generate duplication of the target DNA upon insertion, and (5) excises precisely.
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References
Benton WD, Davis RW (1977) Screening λ gt recombinant clones by hybridization to single plaques in situ. Science 196:180
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
Borderon E, Bieth G, Horodniceanu T (1982) Genetic and physical studies of Streptococcus faecalis hemolysin plasmids. FEMS Microbiol Lett 14:51–55
Boyer HW, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472
Caillaud F, Carlier C, Courvalin P (1987) Physical analysis of the conuugative shuttle transposon Tn1545. Plasmid 17:58–60
Courvalin P, Carlier C (1986) Transposable multiple antibiotic resistance in Streptococcus pneumoniae. Mol Gen Genet 205:291–297
Courvalin P, Carlier C (1987) Tn1545: a conjugative shuttle transposon. Mol Gen Genet 206:259–264
Courvalin P, Carlier C, Collatz E (1980) Plamid-mediated resistance to aminocyclitol antibiotics in group D streptococci. J Bacteriol 143:541–551
Covarrubias L, Bolivar F (1982) Construction and characterisation of new cloning vehicles. VI. Plasmid pBR329, a new derivative of pBR328 lacking the 482 base-pair inverted duplication. Gene 17:79–89
Di Nocera P, Digan M (1983) A family of oligo-adenylate-terminated transposable sequences in Drosophila melanogaster. J Mol Biol 168:715–727
Grindley N, Reed R (1985) Transpositional recombination in prokaryotes. Annu Rev Biochem 54:863–896
Kahman R, Kamp D (1979) Nucleotide sequence of the attachment sites of bacteriophage Mu DNA. Nature 280:247–250
Koo H-S, Wu H-M, Crothers DM (1986) DNA bending at adenine-thymine tracts. Nature 320:501–506
Labigne-Roussel A, Witchitz J, Courvalin P (1982) Modular evolution of disseminated Inc7-M plasmids encoding gentamicin resistance. Plasmid 8:215–231
Lam S, Roth JR (1986) Structural and functional studies of insertion element IS200. J Mol Biol 187:157–167
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Messing J (1979) A multipurpose cloning system based on the single-stranded bacteriophage M13. Recombinant DNA Tech Bull 1:43–44
Murphy E, Löfdahl S (1984) Transposition of Tn554 does not generate a target duplication. Nature 307:292–294
Normore WM (1976) Guanine plus cytosine (GC) composition of the DNA of bacteria, fungi, algae and protozoa. In: Fasman GD (ed) Handbook of biochemistry and molecular biology, vol II. CRC Press, Cleveland, Ohio, pp 65–240
Novick RP, Clowes RC, Cohen SN, Curtiss III R, Datta N, Falkow S (1976) Uniform nomenclature for bacterial plasmids: a proposal. Bacteriol Rev 40:168–189
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Schachman HK, Adler J, Radding CM, Lehman IR, Kornberg A (1960) Enzymatic synthesis of deoxyribonucleic acid. J Biol Chem 235:3242–3249
Schnabel HP, Palm P, Dick K, Gramp B (1984) Sequence analysis of the insertion element ISH1.8 and of associated structural changes in the genome of phage ϕ H of the archaebacterium Halobacterium halobium. EMBO J 3:1717–1722
Shapiro JA, ed (1983) In: Mobile genetic elements. Academic Press, New York
Shinnick TM, Lund E, Smithies O, Blattner FR (1975) Hybridization of labeled RNA to DNA in agarose gels. Nucleic Acids Res 2:1911–1929
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Staden R (1980) A new computer method for storage and manipulation of DNA gel reading data. Nucleic Acids Res 8:3673–3694
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Caillaud, F., Courvalin, P. Nucleotide sequence of the ends of the conjugative shuttle transposon Tn1545 . Mol Gen Genet 209, 110–115 (1987). https://doi.org/10.1007/BF00329844
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DOI: https://doi.org/10.1007/BF00329844