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PMID: 6186991 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Accurate and efficient in vitro splicing of purified precursor RNAs specified by early region 2 of the adenovirus 2 genome.

Nucleic acids research ·Vol. 11 ·No. 5 ·1983-03-11 ·Pages 1337-48

Goldenberg CJ, Hauser SD

Abstract

Polyadenylated and deproteinized nuclear RNA precursors encoded by early region 2 of the adenovirus 2 genome are spliced in vitro by nuclear extracts prepared from MOPC-315 mouse myeloma cells. The in vitro reaction excises sequences from two introns and attaches 5' sequences to the mRNA body. The nucleotide sequence across the splice junctions in the E2 RNAs processed in vitro was investigated by performing primer extensions in the presence of dideoxynucleotides and direct sequencing on polyacrylamide gels. We conclude that the in vitro splicing reaction is accurate and has the same precision as that of in vivo E2 cytoplasmic mRNA prepared from Ad2 infected cells. The efficiency of in vitro splicing by the nuclear extracts is very high. Approximately 80% of E2 RNA precursor, on a molar basis, are spliced in vitro to a mature RNA. These findings provide evidence that a nuclear extract prepared from MOPC-315 mouse myeloma cells is capable of accurate and efficient splicing of E2 RNA precursors.

MeSH Terms
Adenoviruses, Human/genetics Animals Base Sequence Carcinoma Cell Line Cell Nucleus/metabolism Genes Genes, Viral Humans Mice Mouth Neoplasms Nucleic Acid Hybridization Plasmacytoma Poly A/genetics RNA/genetics RNA, Messenger RNA, Viral/genetics Transcription, Genetic
Chemicals
RNA, Messenger RNA, Viral Poly A RNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Goldenberg C J
Hauser S D
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38 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/11.5.1337
Published
1983-03-11
Pages
1337-48
Language
English
Region
England
NLM ID
0411011
PMCID
PMC325800
Subset
IM
Grants
NIAID NIH HHS · IR01 AI 19370-01 · United States
Databases
GENBANK
J01917, J01918, J01919, J01920, J01921, J01922, J01923, J01924, J01925, J01926, J01927, J01928, J01929, J01930, J01931, J01932, J01933, J01934, J01935, J01936, J01937, J01938, J01939, J01940, J01941, J01942, J01943, J01944, J01945, J01946
External Links
PubMed source DOI: 10.1093/nar/11.5.1337 PMC Full Text View journal details
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