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PMID: 6589618 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

In vitro-synthesized adenovirus 2 messenger RNA precursors are accurately spliced by nuclear extracts.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 81 ·No. 15 ·1984-08-00 ·Pages 4707-11

Goldenberg CJ

Abstract

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.

MeSH Terms
Adenoviruses, Human/genetics Animals Cell Nucleus/ultrastructure Cell-Free System Cells, Cultured Kinetics Mice Nucleic Acid Precursors/genetics RNA Processing, Post-Transcriptional RNA Splicing RNA, Messenger/genetics RNA, Viral/genetics
Chemicals
Nucleic Acid Precursors RNA, Messenger RNA, Viral
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Goldenberg C J
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26 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.81.15.4707
Published
1984-08-00
Pages
4707-11
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC391559
Subset
IM
Grants
NIAID NIH HHS · AI-19370 · United States
External Links
PubMed source DOI: 10.1073/pnas.81.15.4707 PMC Full Text View journal details
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