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- W1976220310 abstract "Nested deletions from one end of the genomic DNA in bacterial artificial chromosomes (BACs) and P1 artificial chromosomes (PACs) are readily generated by inserting a loxP site-containing Tn10 minitransposon into the recombinant clone and transducing with P1 phage. Although the size of clones in the deletion series is largely random, in about 5% of BACs and PACs the distribution appears skewed to a certain length, and in rare cases (<1%) is definitely skewed to a particular size. Here we investigate this relatively rare phenomenon and validate that sequence-specific transposon insertions are not the cause of such skewed nested-deletion libraries. Instead, a detailed analysis of our experiments with a BAC clone demonstrating this unusual feature indicates that deletions of a certain size arise from clonal expansion of a transposon insertion as a result of transient derepression of the transposase gene prior to IPTG induction. Transposition itself shows no bias to any particular region of insert DNA in the clone. We suggest a simple modification to the procedure for generating nested-deletions that allows all BACs and PACs to produce nested-deletions of random size. These findings should provide additional insight into the causes of site selectivity in genomic clones with other inducible transposon systems." @default.
- W1976220310 created "2016-06-24" @default.
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- W1976220310 date "2000-10-01" @default.
- W1976220310 modified "2023-09-26" @default.
- W1976220310 title "Analysis of a Clonal Selection Event during Transposon-Mediated Nested-Deletion Formation in Rare BAC and PAC Clones" @default.
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- W1976220310 doi "https://doi.org/10.1006/abio.2000.4750" @default.
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