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Complete sequence of a mariner transposable element from the predatory mite Metaseiulus occidentalis isolated by an inverse PCR approach
Authors:A. Jeyaprakash  M. A. Hoy
Affiliation:Department of Entomology and Nematology, University of Florida, Gainesville, Florida, USA
Abstract:Degenerate primers designed and synthesized based on two conserved regions of the mariner transposase open reading frame were used to amplify a 454 bp DNA fragment from M. occidentalis. Two inverse primers were then synthesized and used to amplify flanking genomic DNA fragments from M. occidentalis by a ligation-mediated inverse PCR. The complete mariner element (Moc 1) was 1284 bp long, including the imperfect 28 bp inverted terminal repeat sequences, and shared 59% similarity to an active 1286 bp long D. mauritiana mariner element (Mos 1). Insertions, deletions and substitutions were observed in the Moc 1 sequence at several positions. No intact open reading frame was detected and the Moc 1 element is considered inactive. Stringent Southern blot hybridizations revealed at least twelve copies of mariner sequences similar to Moc 1 in the colonies tested.
Keywords:cloning    sequence    inverse    PCR    mariner    transposable    element    Metaseiulus occidentalis    Phytoseiidae.
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