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. 2010 Sep;11(9):728-34.
doi: 10.1631/jzus.B1000139.

Using chimeric piggyBac transposase to achieve directed interplasmid transposition in silkworm Bombyx mori and fruit fly Drosophila cells

Na Wang  1 , Cai-ying Jiang, Ming-xing Jiang, Chuan-xi Zhang, Jia-an Cheng
Affiliations

Using chimeric piggyBac transposase to achieve directed interplasmid transposition in silkworm Bombyx mori and fruit fly Drosophila cells

Na Wang et al. J Zhejiang Univ Sci B. 2010 Sep.

Abstract

The piggyBac transposon has been long used to integrate foreign DNA into insect genomes. However, undesirable transgene expression can result from random insertions into the genome. In this study, the efficiency of chimeric Gal4-piggyBac transposase in directing integration onto a DNA target plasmid was evaluated in cultured silkworm Bombyx mori Bm-12 and fruit fly Drosophila Schneider 2 (S2) cells. The Gal4-piggyBac transposase has a Gal4 DNA-binding domain (DBD), and the target plasmid has upstream activating sequences (UAS) to which the Gal4 DBD can bind with high affinity. The results indicate that, in the Bm-12 and S2 cells, transpositional activity of Gal4-piggyBac transposase was increased by 4.0 and 7.5 times, respectively, compared to controls, where Gal4-UAS interaction was absent. Moreover, the Gal4-piggyBac transposase had the ability of directing piggyBac element integration to certain sites of the target plasmid, although the target-directing specificity was not as high as expected. The chimeric piggyBac transposase has the potential for use in site-directed transgenesis and gene function research in B. mori.

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Figures

Fig. 1
Fig. 1
Insertion sites of piggyBac donor elements in pGDV1-UAS and pGDV1 (control, lacking UAS target site) target plasmids in the transposition assays performed in Bombyx mori Bm-12 cells (a) and Drosophila S2 cells (b) Solid and open circles represent the insertions into the pGDV1-UAS and pGDV1 targets, respectively. The circles below the line represent the insertions occurring in an opposite direction relative to the Cam gene. The numbers represent the nucleotide locations on the pGDV1 plasmid. The solid box stands for the UAS site on the pGDV1-UAS plasmid
Fig. 1
Fig. 1
Insertion sites of piggyBac donor elements in pGDV1-UAS and pGDV1 (control, lacking UAS target site) target plasmids in the transposition assays performed in Bombyx mori Bm-12 cells (a) and Drosophila S2 cells (b) Solid and open circles represent the insertions into the pGDV1-UAS and pGDV1 targets, respectively. The circles below the line represent the insertions occurring in an opposite direction relative to the Cam gene. The numbers represent the nucleotide locations on the pGDV1 plasmid. The solid box stands for the UAS site on the pGDV1-UAS plasmid
See this image and copyright information in PMC

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