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Proc Natl Acad Sci U S A. Feb 15, 1991; 88(4): 1247–1250.
PMCID: PMC50994

Transposon-facilitated DNA sequencing.

Abstract

We describe here a transposon-based DNA sequencing strategy that allows the introduction of sequencing priming sites throughout a target sequence by bacterial mating. A miniplasmid was designed to select against transposon insertions into the vector. Sites of transposon insertion are mapped by the polymerase chain reaction with bacterial overnight cultures providing the templates. A small set of plasmids with transposons spaced several hundred base pairs apart can then be sequenced. Sequencing primers corresponding to the transposon ends allow sequencing in both directions. Thus, the entire sequence of both strands can be easily determined.

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Selected References

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  • Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. [PubMed]
  • Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. [PubMed]
  • Strauss EC, Kobori JA, Siu G, Hood LE. Specific-primer-directed DNA sequencing. Anal Biochem. 1986 Apr;154(1):353–360. [PubMed]
  • Guyer MS. Uses of the transposon gamma delta in the analysis of cloned genes. Methods Enzymol. 1983;101:362–369. [PubMed]
  • Ahmed A. A rapid procedure for DNA sequencing using transposon-promoted deletions in Escherichia coli. Gene. 1985;39(2-3):305–310. [PubMed]
  • Adachi T, Mizuuchi M, Robinson EA, Appella E, O'Dea MH, Gellert M, Mizuuchi K. DNA sequence of the E. coli gyrB gene: application of a new sequencing strategy. Nucleic Acids Res. 1987 Jan 26;15(2):771–784. [PMC free article] [PubMed]
  • Liu L, Whalen W, Das A, Berg CM. Rapid sequencing of cloned DNA using a transposon for bidirectional priming: sequence of the Escherichia coli K-12 avtA gene. Nucleic Acids Res. 1987 Nov 25;15(22):9461–9469. [PMC free article] [PubMed]
  • Nag DK, Huang HV, Berg DE. Bidirectional chain-termination nucleotide sequencing: transposon Tn5seq1 as a mobile source of primer sites. Gene. 1988 Apr 15;64(1):135–145. [PubMed]
  • Barrett BK, Berget PB. Using transposon Tn5 insertions to sequence bacteriophage T4 gene 11. DNA. 1989 May;8(4):287–295. [PubMed]
  • Guyer MS. The gamma delta sequence of F is an insertion sequence. J Mol Biol. 1978 Dec 15;126(3):347–365. [PubMed]
  • Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. [PubMed]
  • Casadaban MJ, Cohen SN. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. [PubMed]
  • Sutcliffe JG. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. [PubMed]
  • Holmes DS, Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. [PubMed]
  • Goto N, Shoji A, Horiuchi S, Nakaya R. Conduction of nonconjugative plasmids by F' lac is not necessarily associated with transposition of the gamma delta sequence. J Bacteriol. 1984 Aug;159(2):590–596. [PMC free article] [PubMed]
  • Palazzolo MJ, Hyde DR, VijayRaghavan K, Mecklenburg K, Benzer S, Meyerowitz E. Use of a new strategy to isolate and characterize 436 Drosophila cDNA clones corresponding to RNAs detected in adult heads but not in early embryos. Neuron. 1989 Oct;3(4):527–539. [PubMed]
  • Strausbaugh LD, Bourke MT, Sommer MT, Coon ME, Berg CM. Probe mapping to facilitate transposon-based DNA sequencing. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6213–6217. [PMC free article] [PubMed]

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