In vitro splicing of SV40 late mRNA in isolated nuclei from CV-1 cells.
AUTOR(ES)
Hamada, H
RESUMO
An in vitro splicing system utilizing isolated nuclei of SV40 infected cells has been developed. Nuclei were isolated from CV-1 cells at a late stage of SV40 infection after a pulse-labeling with 3H-uridine. In nuclei prepared under mild isotonic conditions, 19S viral coded RNA synthesized in vivo was converted in vitro into 16S mRNA. In contrast, the nuclei prepared with RSB, a hypotonic medium, showed a very low splicing activity only. Addition of a "nuclear extract" to these nuclei restored the activity almost to the original level. These results indicate that 1) 19S RNA is indeed a precursor to 16S mRNA 2) the splicing of 19S RNA into 16S RNA takes place in the nucleus, and 3) at least a part of the enzyme system required for splicing could be extracted from the nucleus. This in vitro system may be useful for the assay of the splicing enzyme(s).
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=327292Documentos Relacionados
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