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 Citations (Scopus)

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
Pages (from-to)4215-4220
Number of pages6
JournalBiochemistry
Volume14
Issue number19
StatePublished - Dec 1 1975
Externally publishedYes

Fingerprint

Sepharose
Purification
RNA
Messenger RNA
DNA
Viral Structural Proteins
RNA Viruses
Viruses
Cyanogen Bromide
Complementary RNA
Covalent bonds
Cell Extracts
Triticum

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification of SV-40 messenger RNA by hybridization to SV-40 DNA covalently bound to sepharose. / Gilboa, Eli; Prives, Carol L.; Aviv, Haim.

In: Biochemistry, Vol. 14, No. 19, 01.12.1975, p. 4215-4220.

Research output: Contribution to journalArticle

Gilboa, Eli ; Prives, Carol L. ; Aviv, Haim. / Purification of SV-40 messenger RNA by hybridization to SV-40 DNA covalently bound to sepharose. In: Biochemistry. 1975 ; Vol. 14, No. 19. pp. 4215-4220.
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