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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).
H Hamada, and
T Igarashi, and
M Muramatsu
April 1976,
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H Hamada, and
T Igarashi, and
M Muramatsu
December 1986,
Nucleic acids research,
H Hamada, and
T Igarashi, and
M Muramatsu
October 1978,
Nature,
H Hamada, and
T Igarashi, and
M Muramatsu
March 1977,
Biochemical and biophysical research communications,
H Hamada, and
T Igarashi, and
M Muramatsu
January 1979,
Proceedings of the National Academy of Sciences of the United States of America,
H Hamada, and
T Igarashi, and
M Muramatsu
May 1978,
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H Hamada, and
T Igarashi, and
M Muramatsu
January 1970,
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H Hamada, and
T Igarashi, and
M Muramatsu
January 2000,
Tsitologiia,
H Hamada, and
T Igarashi, and
M Muramatsu
January 1974,
Intervirology,
H Hamada, and
T Igarashi, and
M Muramatsu
February 1975,
Virology,
