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http://dx.doi.org/10.7852/jses.2013.51.2.153

Production of fluorescent green silk using fibroin H-chain expression system  

Kim, Seong Wan (National Academy of Agricultural Science, RDA)
Yun, Eun Young (National Academy of Agricultural Science, RDA)
Choi, Kwang-Ho (National Academy of Agricultural Science, RDA)
Kim, Seong Ryul (National Academy of Agricultural Science, RDA)
Park, Seung Won (Department of Biotechnology, Catholic University of Daegu)
Kang, Seok Woo (National Academy of Agricultural Science, RDA)
Goo, Tae Won (National Academy of Agricultural Science, RDA)
Publication Information
Journal of Sericultural and Entomological Science / v.51, no.2, 2013 , pp. 153-158 More about this Journal
Abstract
To express green fluorescent protein in the cocoon of silkworm, we constructed the fibroin H-chain expression system to produce enhanced green fluorescent protein (EGFP) in the cocoon of transgenic silkworms. The EGFP fusion protein, each with N- and C-terminal sequences of the fibroin H-chain, was designed to be secreted into the lumen of the posterior silk glands. The expression of the EGFP/H-chain fusion gene was regulated by the fibroin H-chain promoter. The use of the 3xP3-driven DsRed2 cDNA as a marker allowed us to rapidly distinguish transgenic silkworm. A mixture of the donor and helper vector was micro-injected into 1,200 eggs of bivoltin silkworms, Baegokjam. We obtained 8 broods. The cocoon displayed strong green fluorescence, proving that the fusion protein was present in the cocoon. Also, the presence of fusion proteins in cocoons was demonstrated by SDS-PAGE and immunoblotting. Accordingly, we suggest that the EGFP fluorescence silk will enable the production of the novel biomaterial based on the transgenic silk.
Keywords
Fluorescent; Transgenic silkworm; EGFP; Bombyx mori; Silk;
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