By Karl Maramorosch, Gordon H. Sato
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Additional resources for Advances in cell culture. Volume 6
A) K14-Transfected mouse 3T3 fibroblast, anti-type I epidermal keratin antibody; (B) K6b-transfected mouse 3T3 fibroblast, anti-type II epidermal keratin antibody; (C) K6b-transfected mouse 3T3 fibroblast, anti-type I epidermal keratin antibody; (D) K6b-transfected marsupial PtK2 epithelial cell, anti-type II epidermal keratin antibody. Note: The anti-type II epidermal keratin antibody does not cross-react with the endogenous PtK2 keratin network. An anti-type I epidermal keratin antibody did not stain the K6b-transfected PtK2 cells.
10. The expression of human epidermal keratin genes in transfected fibroblasts and simple epithelial cells. A human type I epidermal keratin gene encoding K14 was inserted into a bacterial plasmid containing an SV40 enhancer sequence. A promoterless human type II epidermal keratin gene encoding K6b was inserted into a bacterial plasmid containing an SV40 promoter and enhancer sequence. The vectors were then transfected by the calcium phosphate method into fibroblasts or simple epithelial cells.
Some regulatory sequences of abundantly expressed, specialized genes have been found to share homology with a consensus sequence G T G G A A A G (Banerji et al, 1981; Queen and Baltimore, 1983; Rosenthal et al, 1983; Gorski et al, 1986). First identified in SV40, these sequences are thought to be target sites for binding of tissue-specific proteins that can confer increased transcriptional activity to a gene (for review, see Scholer and Gruss, 1984). Consequently, these sequences have been called enhancer elements.