Transposon-facilitated DNA sequencing.
- 15 February 1991
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 88 (4) , 1247-1250
- https://doi.org/10.1073/pnas.88.4.1247
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.Keywords
This publication has 16 references indexed in Scilit:
- Use of a new strategy to isolate and characterize 436 drosophila cDNA clones corresponding to RNAs detected in adult heads but not in early embryosNeuron, 1989
- Using Transposon Tn5 Insertions to Sequence Bacteriophage T4 Gene11DNA, 1989
- Bidirectional chain-termination nucleotide sequencing: transposon Tn5seq1 as a mobile source of primer sitesGene, 1988
- Specific-primer-directed DNA sequencingAnalytical Biochemistry, 1986
- Enzymatic Amplification of β-Globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell AnemiaScience, 1985
- A rapid procedure for DNA sequencing using transposon-promoted deletions in Escherichia coliGene, 1985
- Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencingGene, 1984
- Analysis of gene control signals by DNA fusion and cloning in Escherichia coliJournal of Molecular Biology, 1980
- Complete Nucleotide Sequence of the Escherichia coli Plasmid pBR322Published by Cold Spring Harbor Laboratory ,1979
- The λδ sequence of F is an insertion sequenceJournal of Molecular Biology, 1978