New procedure for isolation of Rous sarcoma virus-specific RNA from infected cells. 1979

P A Bromley, and P F Spahr, and J L Darlix

The use of mercurated "strong stop" complementary DNA (complementary to the 5'-terminal 101 nucleotides of Rous sarcoma virus RNA) in the isolation of virus-specific RNA from infected chicken embryo fibroblasts is described. Strong stop Rous sarcoma virus complementary DNA was mercurated chemically, and, as a result of the low complexity of this DNA, short hybridization times (up to 15 min) and heating in the absence of formamide were found to be adequate conditions for the isolation of virus-specific RNA. The purity of the isolated RNA was demonstrated by analysis of labeled RNase T1-resistant oligonucleotides by two-dimensional polyacrylamide gel electrophoresis. The isolated RNA could be translated in the in vitro protein synthesis system derived from rabbit reticulocytes, and an analysis of polypeptides programmed by isolated RNA before and after immunoprecipitation further demonstrated both the purity of the isolated mRNA and the quantitative nature of the isolation procedure.

UI MeSH Term Description Entries
D008628 Mercury A silver metallic element that exists as a liquid at room temperature. It has the atomic symbol Hg (from hydrargyrum, liquid silver), atomic number 80, and atomic weight 200.59. Mercury is used in many industrial applications and its salts have been employed therapeutically as purgatives, antisyphilitics, disinfectants, and astringents. It can be absorbed through the skin and mucous membranes which leads to MERCURY POISONING. Because of its toxicity, the clinical use of mercury and mercurials is diminishing.
D009189 Avian Myeloblastosis Virus A species of ALPHARETROVIRUS causing anemia in fowl. Myeloblastosis Virus, Avian,Avian Myeloblastosis Viruses,Myeloblastosis Viruses, Avian,Virus, Avian Myeloblastosis,Viruses, Avian Myeloblastosis
D009693 Nucleic Acid Hybridization Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503) Genomic Hybridization,Acid Hybridization, Nucleic,Acid Hybridizations, Nucleic,Genomic Hybridizations,Hybridization, Genomic,Hybridization, Nucleic Acid,Hybridizations, Genomic,Hybridizations, Nucleic Acid,Nucleic Acid Hybridizations
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D002642 Chick Embryo The developmental entity of a fertilized chicken egg (ZYGOTE). The developmental process begins about 24 h before the egg is laid at the BLASTODISC, a small whitish spot on the surface of the EGG YOLK. After 21 days of incubation, the embryo is fully developed before hatching. Embryo, Chick,Chick Embryos,Embryos, Chick
D004259 DNA-Directed DNA Polymerase DNA-dependent DNA polymerases found in bacteria, animal and plant cells. During the replication process, these enzymes catalyze the addition of deoxyribonucleotide residues to the end of a DNA strand in the presence of DNA as template-primer. They also possess exonuclease activity and therefore function in DNA repair. DNA Polymerase,DNA Polymerases,DNA-Dependent DNA Polymerases,DNA Polymerase N3,DNA Dependent DNA Polymerases,DNA Directed DNA Polymerase,DNA Polymerase, DNA-Directed,DNA Polymerases, DNA-Dependent,Polymerase N3, DNA,Polymerase, DNA,Polymerase, DNA-Directed DNA,Polymerases, DNA,Polymerases, DNA-Dependent DNA
D004279 DNA, Viral Deoxyribonucleic acid that makes up the genetic material of viruses. Viral DNA
D005347 Fibroblasts Connective tissue cells which secrete an extracellular matrix rich in collagen and other macromolecules. Fibroblast
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001358 Avian Sarcoma Viruses Group of alpharetroviruses (ALPHARETROVIRUS) producing sarcomata and other tumors in chickens and other fowl and also in pigeons, ducks, and RATS. Avian Sarcoma Virus B77,Chicken Sarcoma Virus B77,Chicken Tumor 1 Virus,Fujinami sarcoma virus,Sarcoma Viruses, Avian,Avian Sarcoma Virus,Fujinami sarcoma viruses,Sarcoma Virus, Avian,Virus, Avian Sarcoma,Viruses, Avian Sarcoma,sarcoma virus, Fujinami,virus, Fujinami sarcoma,viruses, Fujinami sarcoma

Related Publications

P A Bromley, and P F Spahr, and J L Darlix
January 1984, Molekuliarnaia biologiia,
P A Bromley, and P F Spahr, and J L Darlix
March 1971, Biochemical and biophysical research communications,
P A Bromley, and P F Spahr, and J L Darlix
August 1964, Proceedings of the National Academy of Sciences of the United States of America,
P A Bromley, and P F Spahr, and J L Darlix
June 1967, Proceedings of the National Academy of Sciences of the United States of America,
P A Bromley, and P F Spahr, and J L Darlix
November 1977, Virology,
P A Bromley, and P F Spahr, and J L Darlix
June 1974, Journal of virology,
P A Bromley, and P F Spahr, and J L Darlix
October 1974, Science (New York, N.Y.),
P A Bromley, and P F Spahr, and J L Darlix
January 1975, Cold Spring Harbor symposia on quantitative biology,
P A Bromley, and P F Spahr, and J L Darlix
July 1972, Journal of virology,
Copied contents to your clipboard!