Procedure for the preparation of milligram quantities of adenovirus messenger ribonucleic acid
- PMID: 4644629
- PMCID: PMC356592
- DOI: 10.1128/JVI.10.6.1126-1129.1972
Procedure for the preparation of milligram quantities of adenovirus messenger ribonucleic acid
Abstract
Late after adenovirus 2 infection (18 hr), nearly all newly synthesized polysomal messenger ribonucleic acid (mRNA) is viral specified. Large amounts of adenovirus mRNA have been purified by utilizing membrane filtration at high ionic strength. With this procedure, molecules that contain polyadenylic acid [poly (A)] tracts are bound selectively, and ribosomal RNA can be separated from the viral mRNA which contains poly(A). Polysomal RNA synthesized 18 hr after infection was labeled in the presence of 0.02 mug of actinomycin D per ml and extracted at pH 9.0. This RNA annealed 40% to 3 mug of adenovirus 2 deoxyribonucleic acid; the RNA selected by membrane filtration bound 80% under the same conditions. The RNA eluted from membrane filters was 80 to 90% greater than 18S and contained species migrating as 31, 27, and 24S. Binding of polysomal RNA to individual membrane filters was linear, using as much as 300 mug of RNA per membrane. A 1.1-mg amount of viral RNA was prepared from 17.7 mg of polysomal RNA that had been purified by extraction at pH 9.0.
Similar articles
-
Isolation and characterization of adenovirus messenger ribonucleic acid in productive infection.J Virol. 1972 Nov;10(5):909-19. doi: 10.1128/JVI.10.5.909-919.1972. J Virol. 1972. PMID: 4636395 Free PMC article.
-
Isolation of DNA Strand-specific early messenger RNA species in cells infected by human adenovirus 2.Proc Natl Acad Sci U S A. 1974 Aug;71(8):2951-5. doi: 10.1073/pnas.71.8.2951. Proc Natl Acad Sci U S A. 1974. PMID: 4528619 Free PMC article.
-
Localization of adenovirus 2 messenger RNA's to segments of the viral genome defined by endonuclease R-R1.Proc Natl Acad Sci U S A. 1974 Oct;71(10):4057-61. doi: 10.1073/pnas.71.10.4057. Proc Natl Acad Sci U S A. 1974. PMID: 4610567 Free PMC article.
-
Synthesis of virus-specific ribonucleic acid in KB cells infected with type 2 adenovirus.J Virol. 1971 Aug;8(2):203-14. doi: 10.1128/JVI.8.2.203-214.1971. J Virol. 1971. PMID: 4256015 Free PMC article.
-
Two classes of cytoplasmic viral RNA synthesized early in productive infection with adenovirus 2.J Virol. 1974 Oct;14(4):751-7. doi: 10.1128/JVI.14.4.751-757.1974. J Virol. 1974. PMID: 4418388 Free PMC article.
Cited by
-
In vitro synthesis of adenovirus core proteins.J Virol. 1974 Aug;14(2):375-83. doi: 10.1128/JVI.14.2.375-383.1974. J Virol. 1974. PMID: 4211166 Free PMC article.
-
Role of DNA-dependent RNA polymerases II and III in transcription of the adenovirus genome late in productive infection.Proc Natl Acad Sci U S A. 1974 Sep;71(9):3426-39. doi: 10.1073/pnas.71.9.3426. Proc Natl Acad Sci U S A. 1974. PMID: 4530313 Free PMC article.
-
Simple and rapid method for disruption of bacteria for protein studies.Appl Environ Microbiol. 1983 Oct;46(4):941-3. doi: 10.1128/aem.46.4.941-943.1983. Appl Environ Microbiol. 1983. PMID: 6639038 Free PMC article.
-
Identification of early adenovirus type 2 RNA species transcribed from the left-hand end of the genome.J Virol. 1975 Oct;16(4):905-12. doi: 10.1128/JVI.16.4.905-912.1975. J Virol. 1975. PMID: 1165597 Free PMC article.
-
Sequence relationships between adenovirus 2 early RNA and viral RNA size classes synthesized at 18 hours after infection.J Virol. 1975 Jan;15(1):137-44. doi: 10.1128/JVI.15.1.137-144.1975. J Virol. 1975. PMID: 1113370 Free PMC article.
References
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
