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Analysis of transgenic silkworms producing insulin-like growth factor-I in Bombyx mori
Authors:Jiyeon Seong  Min-Jung Kim  Soo Won Lee  Hee Jin Yang  Hong Sik Kong  Keun-Chong Kim  Dong Sang Suh
Institution:1. Department of Genetic Engineering, Sungkyunkwan University, Suwon, 440-746, Republic of Korea
3. Genomic Informatics Center, Hankyong National University, Anseong, 425-839, Republic of Korea
2. Sungkyun Biotech, Co., Ltd., Ansan, 425-839, Republic of Korea
4. Hong-Ik University, Jochiwon, 339-700, Republic of Korea
Abstract:Silkworms contain a powerful and effective fibroin promoter, which controls the expression of fibroin, a silk protein. The fibroin promoter and well-known characteristics of silkworm, the application of transgenic technique to silkworm will provide an excellent opportunity to mass-produce biomolecules. In this study, the production of recombinant human insulin like growth factor-I (rhIGF-I) in the silkworm system was designed. The method makes use of the microinjection technique and P element vector to transfer foreign genes into the chromosomes. We constructed the expression vector using the fibroin gene promoter and P element vector containing IGF-I gene (pFpIGF-I). We then microinjected this vector into eggs, and through PCR screening, transgenic silkworms were selected. We isolated and purified rhIGF-I from silkworm cocoons, returning a concentration of rhIGF-I of about 1,300 ng/g from transgenic silkworm cocoons. In a comparison of transgenic silkworm rhIGF-I and colostral IGF-I on cell proliferation, colostral IGF-I was better able to increase the proliferation rate of the cell line relative to the transgenic silkworm rhIGF-I, and showed a similar cell proliferation pattern. The anti-cancer effects of transgenic silkworm rhIGF-I were higher than that of colostral IGF-I on HeLa and SNU-C1 cancer cells. These results confirmed the construction of new transgenic silkworm strains producing rhIGF-I.
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