Silkworm eggs: An ideal model for studying the biological effects of low energy Ar ion interaction in animals |
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Authors: | Lin LingXuelan Liu Jiaping Xu Zhengying YouJingbo Zhou |
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Affiliation: | School of Life Sciences, Anhui Agricultural University, Hefei 230031, China |
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Abstract: | The object of the current work was to study low energy Ar+ ion beam interactions with silkworm eggs and thus provide further understanding of the mechanisms involved in ion bombardment-induced direct gene transfer into silkworm eggs. In this paper, using low-energy Ar+ ion beam bombardment combined with piggyBac transposon, we developed a novel method to induce gene transfer in silkworm. Using bombardment conditions optimized for egg-incubation (25 keV with ion fluences of 800 × 2.6 × 1015 ions/cm2 in dry state under vacuum), vector pBac{3 × P3-EGFPaf} and helper plasmid pHA3pig were successfully transferred into the silkworm eggs. Our results obtained from by PCR assay and genomic Southern blotting analysis of the G1 generations provide evidence that low-energy ion beam can generate some craters that play a role in acting as pathways of exogenous DNA molecules into silkworm eggs. |
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Keywords: | Low-energy Ar+ ion beam Silkworm eggs piggyBac transposon Transgenic silkworm PCR Inverse PCR Southern blotting analysis |
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