Targeted gene replacement in Drosophila via P element-induced gap repair.
Transposable elements of the P family in Drosophila are thought to transpose by a cut-and-paste process that leaves a double-strand gap. The repair of such gaps resulted in the transfer of up to several kilobase pairs of information from a homologous template sequence to the site of P element excision by a process similar to gene conversion. The template was an in vitro-modified sequence that was tested at various genomic positions. Characterization of 123 conversion tracts provided a detailed description of their length and distribution. Most events were continuous conversion tracts that overlapped the P insertion site without concomitant conversion of the template. The average conversion tract was 1379 base pairs, and the distribution of tract lengths fit a simple model of gap enlargement. The conversion events occurred at sufficiently high frequencies to form the basis of an efficient means of directed gene replacement.
Gloor
G B
GB
Faculty of Medicine, Memorial University of Newfoundland, St. John's, Canada.
Nassif
N A
NA
Johnson-Schlitz
D M
DM
Preston
C R
CR
Engels
W R
WR
D000818:Animals; D001483:Base Sequence; D004260:DNA Repair; D004251:DNA Transposable Elements; D004330:Drosophila / Q000235:genetics*; D005785:Gene Conversion; D008957:Models, Genetic; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods; D013698:Templates, Genetic; D014162:Transfection
D004251:DNA Transposable Elements; D015345:Oligonucleotide Probes