title: Expression of the hIGF-I gene driven by the Fhx/P25 promoter in the silk glands of germline silkworm and transformed BmN cells.
abstract: 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 × 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.
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, 215123, Suzhou, People's Republic of China.
authors.forename: Yanmei
authors.initials: Y
authors.lastname: Li
authors.forename: Guangli
authors.initials: G
authors.lastname: Cao
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.forename: Renyu
authors.initials: R
authors.lastname: Xue
authors.forename: Wenlin
authors.initials: W
authors.lastname: Zhou
authors.forename: Chengliang
authors.initials: C
authors.lastname: Gong
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D012831:Bombyx / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005349:Fibroins / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D007334:Insulin-Like Growth Factor I; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014162:Transfection
chemical_list: D006023:Glycoproteins; C418613:fhx protein, Bombyx mori; D007334:Insulin-Like Growth Factor I; D005349:Fibroins