Production of the melittin antimicrobial peptide in transgenic silkworm
2015
Kim, S.W., National Academy of Agricultural Science, RDA, Wanju, Republic of Korea | Goo, T.W., Dongguk University of Gyeongju, Republic of Korea | Kim, S.R., National Academy of Agricultural Science, RDA, Wanju, Republic of Korea | Park, S.W., Catholic University of Daegu, Daegu, Republic of Korea | Choi, K.H., National Academy of Agricultural Science, RDA, Wanju, Republic of Korea
Melittin is the main component of Bee Venom and has antibacterial activity against several bacteria. To produce the melittin antimicrobial peptide, we constructed transgenic silkworm that expressed melittin gene under the control BmA3 promoter using piggyBac vector. The use of the 3xP3-driven EGFP cDNA as a marker allowed us to rapidly distinguish transgenic silkworm. Mixtures of the donor vector and helper vector were micro-injected into 300 eggs of bivoltin silkworms, Baegokjam. In total, 131 larvae (G0) were hatched and allowed to develop into moths. The resulting G1 generation consisted of 36 broods, and we selected 4 broods containing at least 1 EGFP-positive embryo. The rate of successful transgenesis for the G1 broods was 11%. We identified 12 EGFP-positive G1 moths and these were backcrossed with wild-type moths. With the aim of identifying a melittin as antimicrobial peptide, we investigated the Radical diffusion Assay (RDA) and then demonstrated that melittin possesses high antibacterial activities against gramnegative bacteria.
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