Mariner (Mos1) transposase and genomic integration of foreign gene sequences in Bombyx mori cells.
Widespread occurrence in insects and the capacity to transpose in the absence of host-derived factors means that mariner-like elements are considered to be attractive candidates for the development of a universal insect genetic transformation system. Here we show that the Mos1 mariner element of Drosophila mauritiana is capable of mediating excision and transposition events in a silkmoth (Bombyx mori) derived tissue culture cell line (Bm5 cells). Plasmid rescue assays, in combination with Southern hybridization and polymerase chain reaction (PCR) analyses, confirm that the Mos1 transposase can mediate excision of DNA sequences, inserted between terminally repeated sequences recognized by the transposase, and integration into the chromosomal DNA of the Bm5 cells. In addition to chromosomal integration events, inter- and intraplasmid transposition and target element excision events were also detected. Approximately 50% of the plasmids recovered from plasmid rescue assays were found to contain the 'signature' of Mos1-specific excision and/or integration events, indicating that the mariner transposase functions efficiently in the Bombyx cells. Because mariner-induced excision and integration events are strictly dependent on the presence of a co-transfected Mos1 transposase expression vector, it is clear that the multiple copies of endogenous mariner-like elements (Bmmar1) that exist in the Bombyx genome are neither functional nor do they interfere with the efficiency of the transposition process. Thus, the Mos1 element and, probably, mariner elements, in general, hold great promise for the development of genetic transformation systems for lepidopteran insects.
Wang
W
W
Department of Biochemistry, University of Calgary, Canada.
Swevers
L
L
Iatrou
K
K
D000818:Animals; D012831:Bombyx / Q000235:genetics*; D002478:Cells, Cultured; D004251:DNA Transposable Elements; D004330:Drosophila / Q000235:genetics; D015321:Gene Rearrangement; D005822:Genetic Vectors; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic; D019895:Transposases
D004251:DNA Transposable Elements; D019895:Transposases