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That both transgenes require insertion into the same genomic location is strong evidence for this possibility.
Thus it appears likely that functionally equivalent genes may displace their analogs in operons via insertion into the same position.
We generated transgenic Drosophila lines expressing human Aβ43, Aβ42 or Aβ40, using a site-directed integration strategy to allow transgene insertion into the same genomic locus and therefore ensure equivalent levels of Aβ mRNA expression among the lines, as verified by qRT-PCR (p > 0.05, Fig. 1a).
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Such unique sets compensate for multiple insertions into the same location.
Stowaway MITEs have undergone multiple deletions from and insertions into the same site in β-amylase intron 4 during the history of the tribe.
Electroporation can lead to tandem insertions into the same locus, and vector DNA is often digested by exonucleases, making the cloning of insertion sites problematic (Stanford et al. 2001).
Combined, the phylogenetic distribution, sequence variation, and Entrez database comparisons indicate that a Stowaway-like element has undergone multiple deletions from and insertions into the same site in β-amylase intron 4 during the history of the tribe.
A scenario involving multiple insertions into the same site would require as many as five independent acquisitions, or at least three if nodes with low bootstrap support are discounted.
Since the insertion of IS5 into the same position of the parent genome was difficult experimentally, we constructed a cspC deletion strain.
Further PCR analysis confirmed the invariable insertion of tetO into the same chromosomal locus (cjsa_0191) in all clone SA isolates collected since 2003 (over 100 isolates), all of which are resistant to tetracyclines (minimum inhibitory concentration [MIC] ≥16 µg/ml).
Extreme specificity, involving multiple insertions into the exact same site in the genome, has been demonstrated for a variety of transposons.
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