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KMID : 0613820110210121726
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Journal of Life Science
2011 Volume.21 No. 12 p.1726 ~ p.1731
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Construction of Transgenic Silkworms Expressing Human Stem Cell Factor (hSCF)
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Kim Sung-Wan
Yun Eun-Young
Kim Seong-Ryul
Park Seung-Won
Kang Seok-Woo
Goo Tae-Won
Kwon O-Yu
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Abstract
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Human Stem Cell Factor (hSCF) is a cytokine that binds to the c-Kit receptor and plays an important role in hematopoiesis, spermatogenesis, and melanogenesis. To produce the human Stem Cell Factor (hSCF) recombinant protein, we constructed a germline transgenic silkworm using the piggyback vector. The expression of the hSCF gene was driven by the Drosophila heat shock protein 70 (dHsp70) promoter. 3XP3 promotor-driven EGFP was used as a marker which allowed us to rapidly distinguish the transgenic silkworm. A mixture of the donor and helper vector was micro-injected into 1,020 eggs of bivoltin silkworms, Keomokjam. We obtained approximately 22 G1 broods that were EGFP-positive. The expression of the hSCF gene in the transgenic silkworm was analyzed by SDS-PAGE and immunoblotting. Also, analysis of insertion sites into the silkworm genome using inverse PCR showed that exogenous DNA was inserted into the transgenic silkworm genome. These results show that successfully constructed transgenic silkworm expresses the hSCF recombinant protein.
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KEYWORD
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Stem cell factor (SCF), transgenic silkworm, piggyback, EGFP, Bombyx mori
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