New way to isolate simian virus 40 nucleoprotein complexes from infected cells: use of a thiol-specific reagent.

AUTOR(ES)

Boyce, F M

RESUMO

A new method for the isolation of simian virus 40 nucleoprotein complexes from nuclei of lytically infected cells is described. The method is based on the addition of a thiol-specific reagent, 5'5'-dithiobis(2-nitrobenzoic acid), to lysis and extraction buffers. By inhibiting an uncoating activity during simian virus 40 extraction, 5'5'-dithiobis (2-nitrobenzoic acid) allows the use of efficient extraction buffers, such as one containing Triton X-100 and EDTA, for the isolation of native simian virus 40 minichromosomes and virion-type structures. Use of the method is illustrated by following encapsidation of simian virus 40 minichromosomes in a pulse-chase experiment. Since 5'5'-dithiobis (2-nitrobenzoic acid) is an inhibitor of many different enzymes, the 5',5'-dithiobis (2-nitrobenzoic acid) extraction technique may be useful for the isolation of not only papovaviruses but also other viruses and possibly cellular chromatin.

Documentos Relacionados

• Cloning and sequencing of thiol-specific antioxidant from mammalian brain: alkyl hydroperoxide reductase and thiol-specific antioxidant define a large family of antioxidant enzymes.
• Nucleoprotein Complexes in Simian Virus 40-Infected Cells
• Dimerization of thiol-specific antioxidant and the essential role of cysteine 47.
• Association of Simian Virus 40 T Antigen with Simian Virus 40 Nucleoprotein Complexes
• Transcription of reconstituted simian virus 40 nucleoprotein complexes.