By Michael G. Roth (Eds.)
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Extra info for Protein Expression in Animal Cells
And Roth, M. G. (1991). A single amino acid change in the cytoplasmic domain alters the polarized delivery of influenza virus hemagglutinin. J . Cell Biol. 114, 413-421. Bukrinskaya, A. , Vorkunova, N. , Kornliayeva, G. , Narmanbetova, R. , and Vorkunova, G. K. (1982). Influenza virus uncoating in infected cells and effect of rimantadine. J . Gen. Virol. 60,49-59. Bukrinsky, M. , McDonald, T. , Tarpley, W. , andstevenson, M. (1993). Association of integral, matrix, and reverse transcriptase antigens of human immunodeficiency virus type 1 with viral nucleic acids following infection.
3. Following the l-hr adsorption period, the cells are overlaid with growth medium containing 10% NCS and returned to the incubator for 48 hr. 4. The cells are scraped and virus is harvested 48 hr postinfection by three cycles of freezekhawing, followed by centrifugation of cell debris at 2000 rpm for 10 min at 4°C. 5. Supernatants from the centrifugation are stored at - 20°C to - 80°C. Virus titer is determined by plaque assay (see next procedure). b. Plaque Assay We routinely use HEp-2 cells for plaque assay of poliovirus.
2. Remove the 1 :1 mixture and replace with 1 ml of complete Low Viscosity media for 2-4 hr. 3. Prepare three gelatin capsules (size “0”) for each sample; use only the larger portion of each capsule. 4. Fill capsules with fresh Low Viscosity media and place several strips from each sample and then a label into each of the three capsules. 5. Polymerize the resin at 70°C for 24 to 48 hr. Do not cover capsules. 1. Viruses as Model Systems in Cell Biology 35 Acknowledgments We thank Drs. Judith A. Ball, Miroslav Novak, Dan R.
Protein Expression in Animal Cells by Michael G. Roth (Eds.)