Purification of SV-40 Messenger RNA by Hybridization to SV-40 DNA Covalently Bound to Sepharose

Eli Gilboa, Carol L. Prives, Haim Aviv

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

SV-40 DNA sheared form was coupled in a stable covalent bond to cyanogen bromide activated Sepharose. Under the conditions used at least 80% of the DNA was bound to Sepharose. The T1/2 of hybridization of 0.5 μg/ml of SV-40 cRNA to SV-40 DNA-Sepharose was 1 hr. This rate of hybridization is sufficiently rapid to purify SV-40 sequences from solutions containing as little as 0.05-0.1 μg/ml. Nonspecific hybridization of RNA is in the range of 0.1-0.2% of the total input RNA. The DNA-Sepharose is fairly stable and can be reused several times to purify RNA. The SV-40 DNA-Sepharose was used to select large quantities of virus specific RNA from SV-40 infected BS-C-1 cells. The virus specific RNA when added to cell-free extracts from wheat germ was shown to direct the synthesis of the major viral structural protein VP-1.

Original languageEnglish (US)
Pages (from-to)4215-4220
Number of pages6
JournalBiochemistry
Volume14
Issue number19
DOIs
StatePublished - Sep 1 1975
Externally publishedYes

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ASJC Scopus subject areas

  • Biochemistry

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