Transposon-facilitated DNA sequencing.
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.
Strathmann
M
M
California Institute of Technology, Division of Biology, Pasadena 91125.
Hamilton
B A
BA
Mayeda
C A
CA
Simon
M I
MI
Meyerowitz
E M
EM
Palazzolo
M J
MJ
D001483:Base Sequence; D003433:Crosses, Genetic; D004247:DNA / Q000235:genetics*; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000379:methods*; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods; D015183:Restriction Mapping
D004251:DNA Transposable Elements; D015345:Oligonucleotide Probes; D004247:DNA