1. Transgenic silkworms produce recombinant human type III procollagen in cocoons.
- Author
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Tomita, Masahiro, Munetsuna, Hiroto, Sato, Tsutomu, Adachi, Takahiro, Hino, Rika, Hayashi, Masahiro, Shimizu, Katsuhiko, Nakamura, Namiko, Tamura, Toshiki, and Yoshizato, Katsutoshi
- Subjects
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SILKWORMS , *TRANSGENIC organisms , *COLLAGEN - Abstract
We describe the generation of transgenic silkworms that produce cocoons containing recombinant human collagen. A fusion cDNA was constructed encoding a protein that incorporated a human type III procollagen mini-chain with C-propeptide deleted, a fibroin light chain (L-chain), and an enhanced green fluorescent protein (EGFP). This cDNA was ligated downstream of the fibroin L-chain promoter and inserted into a piggyBac vector. Silkworm eggs were injected with the vectors, producing worms displaying EGFP fluorescence in their silk glands. The cocoons emitted EGFP fluorescence, indicating that the promoter and fibroin L-chain cDNAs directed the synthesized products to be secreted into cocoons. The presence of fusion proteins in cocoons was demonstrated by immunoblotting, collagenase-sensitivity tests, and amino acid sequencing. The fusion proteins from cocoons were purified to a single electrophoretic band. This study demonstrates the viability of transgenic silkworms as a tool for producing useful proteins in bulk. [ABSTRACT FROM AUTHOR]
- Published
- 2003
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