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piggyBac ‐mediated somatic transformation of the two‐spotted cricket, Gryllus bimaculatus
Author(s) -
Shinmyo Yohei,
Mito Taro,
Matsushita Takashi,
Sarashina Isao,
Miyawaki Katsuyuki,
Ohuchi Hideyo,
Noji Sumihare
Publication year - 2004
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/j.1440-169x.2004.00751.x
Subject(s) - gryllus bimaculatus , biology , transgene , transposase , somatic cell , gene , transformation (genetics) , transposable element , rna , microbiology and biotechnology , genetics , cricket , genome , ecology
Transgenic insects have been artificially produced to study functions of interesting developmental genes, using insect transposons such as piggyBac . In the case of the cricket, however, transgenic animals have not yet been successfully artificially produced. In the present study, we examined whether the piggyBac transposon functions as a tool for gene delivery in embryos of Gryllus bimaculatus . We used either a piggyBac helper plasmid or a helper RNA synthesized in vitro as a transposase source. An excision assay revealed that the helper RNA was more effective in early Gryllus eggs to transpose a marker gene of eGFP than the helper plasmid containing the piggyBac transposase gene driven by the G. bimaculatus actin3/4 promoter. Further, only when the helper RNA was used, somatic transformation of the embryo with the eGFP gene was observed. These results suggest that the piggyBac system with the helper RNA may be effective for making transgenic crickets.