PARTIAL-PURIFICATION AND PROPERTIES OF A PRE-MESSENGER RNA SPLICING ACTIVITY
- 1 January 1985
- journal article
- research article
- Vol. 260 (2), 1096-1102
Abstract
Precursor RNA substrates for splicing reaction were synthesized in vitro from a plasmid DNA in which the early region 2 gene of adenovirus 2 was fused to an efficient bacteriophage promoter (Salmonella phage 6). Pre-mRNA splicing activity from nuclear extracts of MOPC-315 mouse myeloma cells was partially purified 108-fold by 3 chromatographic steps. The in vitro splicing reaction catalyzed by the partially purified fractions was efficient (60-80% substrate conversion) and accurate at the nucleotide level. The reaction occurred with crude or purified fractions without any detectable lag and nucleotides (ATP or GTP) were absolutely required. Monoclonal anti-Sm antibodies that quantitatively immunoprecipitate U1 small nuclear ribonucleoprotein [RNP] particles totally inhibited the splicing activity of the purified fractions, indicating that U1 small nuclear RNP had co-purified with the activity and were absolutely required for the splicing reaction.This publication has 28 references indexed in Scilit:
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