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Nat Biotechnol. 2000 Jan;18(1):81-4.

Germline transformation of the silkworm Bombyx mori L. using a piggyBac transposon-derived vector.

Author information

  • 1National Institute of Sericultural and Entomological Science, Tsukuba, Ibaraki 305-8634, Japan.

Erratum in

  • Nat Biotechnol 2000 May;18(5):559. Toshiki, T [corrected to Tamura, T]; Chantal, T [corrected to Thibert, C]; Corinne, R [corrected to Royer, C]; Toshio, K [corrected to Kanda, T]; Eappen, A [corrected to Abraham, E]; Mari, K [corrected to Kamba, M]; Natuo, K [corrected to Komoto, N]; Jean-Luc, T [corrected to Thomas, JL]; Bernard, M [corrected to Mauchamp, B]; GĂ©rard, C [corrected to Chavancy, G]; Paul, S [corrected to Shirk, P]; Malcolm, F [corrected to Fraser, M]; Jean-Claude, P [corrected to Prudhomme, JC]; Pierre, C [corrected to Couble, P].

Abstract

We have developed a system for stable germline transformation in the silkworm Bombyx mori L. using piggyBac, a transposon discovered in the lepidopteran Trichoplusia ni. The transformation constructs consist of the piggyBac inverted terminal repeats flanking a fusion of the B. mori cytoplasmic actin gene BmA3 promoter and the green fluorescent protein (GFP). A nonautonomous helper plasmid encodes the piggyBac transposase. The reporter gene construct was coinjected into preblastoderm eggs of two strains of B. mori. Approximately 2% of the individuals in the G1 broods expressed GFP. DNA analyses of GFP-positive G1 silkworms revealed that multiple independent insertions occurred frequently. The transgene was stably transferred to the next generation through normal Mendelian inheritance. The presence of the inverted terminal repeats of piggyBac and the characteristic TTAA sequence at the borders of all the analyzed inserts confirmed that transformation resulted from precise transposition events. This efficient method of stable gene transfer in a lepidopteran insect opens the way for promising basic research and biotechnological applications.

PMID:
10625397
[PubMed - indexed for MEDLINE]
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