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10. Transgenic silkworms produce recombinant human type III procollagen in cocoons.

Author

Tomita, Masahiro, Munetsuna, Hiroto, Sato, Tsutomu, Adachi, Takahiro, Hino, Rika, Hayashi, Masahiro, Shimizu, Katsuhiko, Nakamura, Namiko, Tamura, Toshiki, and Yoshizato, Katsutoshi

Subjects
*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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