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Transgenic silkworms produce recombinant human type III procollagen in cocoons.
- Source :
-
Nature Biotechnology . Jan2003, Vol. 21 Issue 1, p52. 5p. - Publication Year :
- 2003
-
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]
- Subjects :
- *SILKWORMS
*TRANSGENIC organisms
*COLLAGEN
Subjects
Details
- Language :
- English
- ISSN :
- 10870156
- Volume :
- 21
- Issue :
- 1
- Database :
- Academic Search Index
- Journal :
- Nature Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 8820122
- Full Text :
- https://doi.org/10.1038/nbt771