By R. E. Spier
This e-book offers a cutting-edge document at the box of animal mobile know-how, a resource of reference for all these inquisitive about the creation and use of animal telephone items
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USA 77, 4216-4220 Graham and van der Eb (1974): Virology 52, 456 Byrn, R. et al. M. (1980) Biologicals 18, 159-154 Finn, G. K. et al. (1989): Mol. Cell. Biol. 9, 3377-3384 Zastrow, G. et al (1989): Nucl. Acids Research 17,5, 1867-1979 Wegner, M. et al. (1989): Nucl. G. (1991): J. Biol. Chemistry 266, 6000-6005 Schweizer, E. C. et al. (1969): J. Mol. Biol. W. and Wu, R. (1979: Plasmid 2, 536-554 Zheng, H. H. (1990): Nature 344, 170-173 Wurm, F. M. et al. K. ABSTRACT An expression system based on amplification of the glutamine synthetase (gs) gene leads to high level expression of heterologous proteins in CHO and NSO cells.
SFME cell were grown in a one to one mixture of Dulbecco's modified Eagle's medium (DMQO and Ham's F12 (FD medium) supplemented with insulin (10 A/g/ml), transferrin (10 //g/ml), human high density lipoprotein (HDL) (10 //g/ml), and epidermal growth factor (EGF) (50 ng/ml) in dishes precoated with bovine fibronectin (10 μ g/ml). BHK-21 cells were grown in DM04 supplemented with 10% fetal 54 Effector plasmid Reporter plasmid Oncogene« Oncogene (d) Humia c-Ha-rcu fb) Arian ψ-myb 4■ ■ 4 Xbal ■ ■4 Figure 1.
T 100 T r cl. 1 cl. , , • · · • »· ·. · 3 «M» 1 •f- •• • · c-myc-SFME cl. 2 cl. , •. • 9 ** 3« cl. 1 v-myb-SFME cl. 2 cl. , 200 me c/os-SFME cl. 2 cl. 3 tftA * cl. 1 *. ·· . · c-Ha-ras -SFME cl. 2 cl. 3 * ; · . · , · • , 1 , 1 . 1 - 1 L I . Figure 2. Appearance of highly IL-6 productive clones by transfection of oncogenetransformed SFME cells with the IL-6 gene. After transfection of SFME cells (1 x 10 6 cells/ 90mm dish) with 30 //gof ef fector plasmids containing the CMV promoter and oncogenes by high efficiency calcium phos phate coprecipitation method, cells were were selected with 600// g/ml of G418 in 90mm plas tic dishes.