In vitro-synthesized adenovirus 2 messenger RNA precursors are accurately spliced by nuclear extracts.
Precursor mRNAs were synthesized in vitro from a plasmid in which the early region 2 gene of adenovirus 2 is fused to an efficient bacteriophage promoter (Salmonella phage 6). The RNAs were purified and used as substrates for in vitro splicing in the presence of nuclear extracts prepared from MOPC-315 mouse myeloma cells. The in vitro splicing was accurate at the nucleotide level. The reaction occurs rapidly and without any detectable lag. The concentration of the pre-mRNA precursor during incubation appears to be an important factor for high efficiency (60%-80%) of in vitro RNA splicing. Fractionation of the splicing components as well as modifications of the DNA template to study the nucleotide-sequence requirement for in vitro splicing can now be accomplished with this system.
D000260:Adenoviruses, Human / Q000235:genetics*; D000818:Animals; D002467:Cell Nucleus / Q000648:ultrastructure; D002474:Cell-Free System; D002478:Cells, Cultured; D007700:Kinetics; D051379:Mice; D009694:Nucleic Acid Precursors / Q000235:genetics; D012323:RNA Processing, Post-Transcriptional; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics*
D009694:Nucleic Acid Precursors; D012333:RNA, Messenger; D012367:RNA, Viral