Genomic analysis of insertion behavior and target specificity of mini-Tn7 and Tn3 transposons in Saccharomyces cerevisiae.
Transposons are widely employed as tools for gene disruption. Ideally, they should display unbiased insertion behavior, and incorporate readily into any genomic DNA to which they are exposed. However, many transposons preferentially insert at specific nucleotide sequences. It is unclear to what extent such bias affects their usefulness as mutagenesis tools. Here, we examine insertion site specificity and global insertion behavior of two mini-transposons previously used for large-scale gene disruption in Saccharomyces cerevisiae: Tn3 and Tn7. Using an expanded set of insertion data, we confirm that Tn3 displays marked preference for the AT-rich 5 bp consensus site TA[A/T]TA, whereas Tn7 displays negligible target site preference. On a genome level, both transposons display marked non-uniform insertion behavior: certain sites are targeted far more often than expected, and both distributions depart drastically from Poisson. Thus, to compare their insertion behavior on a genome level, we developed a windowed Kolmogorov-Smirnov (K-S) test to analyze transposon insertion distributions in sequence windows of various sizes. We find that when scored in large windows (>300 bp), both Tn3 and Tn7 distributions appear uniform, whereas in smaller windows, Tn7 appears uniform while Tn3 does not. Thus, both transposons are effective tools for gene disruption, but Tn7 does so with less duplication and a more uniform distribution, better approximating the behavior of the ideal transposon.
Seringhaus
Michael
M
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520, USA.
Kumar
Anuj
A
Hartigan
John
J
Snyder
Michael
M
Gerstein
Mark
M
D001483:Base Sequence; D015825:Chromosomes, Fungal / Q000737:chemistry; D016384:Consensus Sequence; D004251:DNA Transposable Elements; D018390:Gene Targeting; D016681:Genome, Fungal; D015698:Genomic Library; D023281:Genomics; D016254:Mutagenesis, Insertional; D016012:Poisson Distribution; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D004251:DNA Transposable Elements