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KMID : 1041220130510020153
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Journal of Sericultural and Entomological Science
2013 Volume.51 No. 2 p.153 ~ p.158
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Production of fluorescent green silk using fibroin H-chain expression system
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Kim Seong-Wan
Yun Eun-Young
Choi Kwang-Ho
Kim Seong-Ryul
Park Seung-Won
Kang Seok-Woo
Goo Tae-Won
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Abstract
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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.
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KEYWORD
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Fluorescent, Transgenic silkworm, EGFP, Bombyx mori, Silk
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