Expression of Inhibitory Proteins
Note: not infrequently over expression of proteins (particularly glycoproteins) is inhibitory to host cells and viral vectors cannot be constructed or propagated. The purpose of the system described here is to inhibit expression by rescuing and growing the desired vectors containing cassettes with expression controlled by a Lac operator, in 293 cells expressing the lac repressor. The vectors can then be used to infect cells lacking the repressor and protein expression at high levels will result. The system is illustrated here.
For more detailed information see reference below and contact DMKS Research (DMKS_Ltd@shaw.ca).
1. McDermott, M.R., Graham, F.L., Hanke, T. and Johnson, D.C Protection of mice against lethal challenge with herpes simplex virus by vaccination with an adenovirus vector expressing HSV glycoprotein B. Virology 169: 244-247, 1989
2. Prevec, L., Schneider, M., Rosenthal, K.L., Belbeck, L.W., Derbyshire, J.B. and Graham, F.L. Use of human adenovirus based vectors for antigen expression in animals. . J. Gen. Virol. 70: 429-434, 1989
References
3. Witmer, L.A., K,L. Rosenthal, F.L. Graham, H.M. Friedman, A. Yee and D.C. Johnson Cytotoxic T lymphocytes specific for herpes simplex virus (HSV) studied using adenovirus vectors expressing HSV glycoproteins J. Gen. Virol. 71: 387-396, 1990
4. Graham, F.L Adenoviruses as expression vectors and recombinant vaccines. Trends in Biotechnology 8: 85-87, 1990
5. Prevec, L., Campbell, J.B., Christie, B.S., Belbeck, L. and Graham, F.L A recombinant human adenovirus vaccine against rabies. J. Inf. Dis. 161: 27-30, 1990
6. Prevec, L. Christie, B.S., Laurie, K.E., Smith, M., Graham, F.L. and Rosenthal, K.L . Immune response to HIV-1 gag antigens induced by recombinant adenovirus vectors in mice and Rhesus macaque monkeys. J. Acquired Imm. Def. Syndr. 4: 568-576, 1991
7. Ebata, S. N., Prevec, L., Graham, F.L., and Dimock, K. Function and Immunogenicity of Human Parainfluenza Virus-3 Glycoproteins Expressed by Recombinant Adenoviruses. Virus Research 24: 21-33, 1992
8. Matthews, D. A., Cummings, D., Evelegh, C., Graham, F. L. and Prevec, L. Development of a modified 293 cell line expressing the lac repressor and its use in the rescue of recombinant adenoviruses expressing high levels of rabies glycoprotein (G). J. Gen. Virol. 80: 345-53, 1999