Journal
BIOTECHNOLOGY LETTERS
Volume 33, Issue 3, Pages 489-494Publisher
SPRINGER
DOI: 10.1007/s10529-010-0462-y
Keywords
BmN cell; Bombyx mori; Fhx/P25 promoter; hIGF-I; Silk gland; Transgenic silkworm
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Funding
- National Basic Research Program of China (973 Program) [2005CB121000]
- National High Technology Research and Development Program of China (863 Program) [2006AA10A119]
- National Natural Science Foundation of China [30571404, 30671590, 31072085]
- Soochow University [Q3134991]
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The expression of the human insulin-like growth factor (hIGF-I) gene driven by the Fhx/P25 promoter in the silk glands of transgenic silkworms (Bombyx mori) and in transformed silkworm cells, was achieved using BmN cells transfected with a piggyBac vector, pigA3GFP-Fhx/P25-hIGF-ie-neo containing a neomycin-resistance gene (neo), a green fluorescent protein gene (gfp), an hIGF-I gene, and a helper plasmid containing the piggyBac transposase sequence under the control of the B. mori actin 3 (A3) promoter. We selected stably transformed BmN cells expressing hIGF-I using the antibiotic G418. The expression level of hIGF-I was about 450 pg in 3 x 10(6) cells, determined by ELISA. The piggyBac vector was transferred into the silkworm eggs using sperm-mediated gene transfer. The expression level of hIGF-I per gram fresh posterior silk glands of G4 transgenic silkworms was approx. 150 ng.
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