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Complete sequence of a mariner transposable element from the predatory mite Metaseiulus occidentalis isolated by an inverse PCR approach
Author(s) -
Jeyaprakash A.,
Hoy M. A.
Publication year - 1995
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/j.1365-2583.1995.tb00005.x
Subject(s) - transposase , transposable element , biology , inverse polymerase chain reaction , open reading frame , genetics , southern blot , genomic dna , inverted repeat , microbiology and biotechnology , insertion sequence , genome , dna , polymerase chain reaction , gene , peptide sequence , multiplex polymerase chain reaction
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.