Production of large amounts of 35S RNA and complementary DNA from avian RNA tumor viruses

R. E. Smith; S. Nebes; J. Leis

doi:10.1016/0003-2697(77)90308-6

Production of large amounts of 35S RNA and complementary DNA from avian RNA tumor viruses

R. E. Smith^*, S. Nebes, J. Leis

^*Corresponding author for this work

Microbiology-Immunology

Research output: Contribution to journal › Article › peer-review

Abstract

An automatic perfusion device was used to collect virus-containing supernatant fluids from transformed and untransformed chick embryo fibroblasts every 2 hr. A 30-day perfusion of 10 roller culture bottles of Rous sarcoma virus-infected cells yielded 520 mg of virus and 18 × 10⁹ transformed cells. Quantitative recovery of 60S and 35S RNA required proteinase K treatment prior to phenol deproteinization. Large amounts of complementary DNA were prepared from a nontransforming virus, derived from avian myeloblastosis virus, by using endogenous reverse transcriptase and proteinase K digestion.

Original language	English (US)
Pages (from-to)	226-234
Number of pages	9
Journal	Analytical Biochemistry
Volume	77
Issue number	1
DOIs	https://doi.org/10.1016/0003-2697(77)90308-6
State	Published - Jan 1977

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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title = "Production of large amounts of 35S RNA and complementary DNA from avian RNA tumor viruses",

abstract = "An automatic perfusion device was used to collect virus-containing supernatant fluids from transformed and untransformed chick embryo fibroblasts every 2 hr. A 30-day perfusion of 10 roller culture bottles of Rous sarcoma virus-infected cells yielded 520 mg of virus and 18 × 109 transformed cells. Quantitative recovery of 60S and 35S RNA required proteinase K treatment prior to phenol deproteinization. Large amounts of complementary DNA were prepared from a nontransforming virus, derived from avian myeloblastosis virus, by using endogenous reverse transcriptase and proteinase K digestion.",

author = "Smith, {R. E.} and S. Nebes and J. Leis",

note = "Funding Information: The authors wish to acknowledge the competent technical assistance of Kay Izard. We wish to thank Bellco Glass, Inc., for their cooperation and skill in the construction of the Autoharvester. This work was supported by NIH Grants No. CA 12323 and CA 14787. J. Leis is the recipient of Research Career Development Award CA 70876.",

year = "1977",

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doi = "10.1016/0003-2697(77)90308-6",

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T1 - Production of large amounts of 35S RNA and complementary DNA from avian RNA tumor viruses

AU - Smith, R. E.

AU - Nebes, S.

AU - Leis, J.

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PY - 1977/1

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Production of large amounts of 35S RNA and complementary DNA from avian RNA tumor viruses

Abstract

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