Animal Virology by David Baltimore (Eds.)

By David Baltimore (Eds.)

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Gen. Virol. 3, 379. , and Monod, J. (1961). J. Mol. Biol. 3, 318. , and Ptashne, M. (1974). Cold Spring Harbor Symp. Quant. Biol. 38, 857. 35 IMMUNOPRECIPITATION OF MURINE LEUKEMIA VIRUSSPECIFIC POLYRIBOSOMES: IDENTIFICATION O F VIRUS-SPECIFIC MESSENGER RNA Nikolaus Mueller -Lantzsch, L o r e n H a u e n and Hung F a n The Salk Institute for Biological Studies San Diego, California 92112 ABSTRACT. An a n t i s e r u m monospecific for the i n t e r n a l s t r u c t u r a l protein p30 of Moloney m u r i n e l e u k e m i a v i r u s (M-MuLV) was p r e p a r e d and used to immunoprecipitate the polyribosomes synthesizing this protein from producer c e l l s .

I. Ara-C (40 μ g/ml final) or chloroquine (50 μ g/ ml final) were added to the culture medium. i. , Hirt extracts (19) were prepared and TCA precipitable material was taken as a measure of viral DNA synthesis ; at the same time the nuclei of the unlabelled cultures were lysed with sarkosyl and RNA polymerase activity of the viral complex was measured by H-UNP incorporation. The rate of viral DNA synthesis was measured in Hirt extracts (19) after a one hour pulse with H-TdR. The viral transcription complex was isolated and assayed in sarkosyl supernatants prepared from unlabelled parallel plates, at the end of the pulse period.

Gielkens, M. H. L . Salden and H . Bloemendal, P r o c . Nat. Acad. Sei. 7 1 , 1093 (1974). S. Jons son and G. Kronvall, E u r . J . Immunol. 4, 29 (1974). P . J . McConahey and F . J . Dixon, Int. A r c h . Allergy 29, 185 (1966). A. P a n e t , D. Baltimore and T. Hanafusa, J . V i r o l . 16, 146 (1975). G. Shanmugam, M. Bhaduri and M. G r e e n , Biochem. Biophys. R e s . C o m m u n . 56, 697 (1974). D . J . Shapiro, J . M. T a y l o r , G. S. McKnight, R. P a l a c i o s , C. Gonzalez, M.

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