Biomaterial Database

Synthesis of simian rotavirus SA11 double-stranded RNA in a cell-free system. 1986

J T Patton

A cell-free system was developed to study the replication of simian rotavirus SA11. The components of the system included (i) subviral particles prepared from infected cells to template the synthesis of viral RNA and (ii) an mRNA-dependent rabbit reticulocyte lysate to support protein synthesis. Based upon nuclease-sensitivity, approximately 20% of the RNA made in vitro was double-stranded (dsRNA) and 80% single-stranded (ssRNA). Electrophoretic analysis of the RNA products on polyacrylamide and low pH agarose gels showed that the system supported the synthesis of 11 dsRNAs and 11 positive-sense ssRNAs that corresponded in size to authentic viral RNAs. The synthesis of dsRNA in vitro was determined to be an asymmetrical process in which a nuclease-sensitive positive-strand RNA acted as a template for the synthesis of negative-strand RNA. The system also supported the initiation of negative-strand RNA using exogenous viral positive-strand RNA as a template. Finally, analysis of subviral particles recovered from reactions suggested that viral proteins made in vitro assembled into nucleoprotein complexes which were similar to those present in infected cells. Together, these results indicate that the cell-free system supported rotavirus RNA replication, transcription and the assembly of subviral particles.

UI	MeSH Term	Description	Entries
D007700	Kinetics	The rate dynamics in chemical or physical systems.
D002460	Cell Line	Established cell cultures that have the potential to propagate indefinitely.	Cell Lines,Line, Cell,Lines, Cell
D002474	Cell-Free System	A fractionated cell extract that maintains a biological function. A subcellular fraction isolated by ultracentrifugation or other separation techniques must first be isolated so that a process can be studied free from all of the complex side reactions that occur in a cell. The cell-free system is therefore widely used in cell biology. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p166)	Cellfree System,Cell Free System,Cell-Free Systems,Cellfree Systems,System, Cell-Free,System, Cellfree,Systems, Cell-Free,Systems, Cellfree
D004587	Electrophoresis, Agar Gel	Electrophoresis in which agar or agarose gel is used as the diffusion medium.	Electrophoresis, Agarose Gel,Agar Gel Electrophoresis,Agarose Gel Electrophoresis,Gel Electrophoresis, Agar,Gel Electrophoresis, Agarose
D004591	Electrophoresis, Polyacrylamide Gel	Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.	Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
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
D012313	RNA	A polynucleotide consisting essentially of chains with a repeating backbone of phosphate and ribose units to which nitrogenous bases are attached. RNA is unique among biological macromolecules in that it can encode genetic information, serve as an abundant structural component of cells, and also possesses catalytic activity. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)	RNA, Non-Polyadenylated,Ribonucleic Acid,Gene Products, RNA,Non-Polyadenylated RNA,Acid, Ribonucleic,Non Polyadenylated RNA,RNA Gene Products,RNA, Non Polyadenylated
D012330	RNA, Double-Stranded	RNA consisting of two strands as opposed to the more prevalent single-stranded RNA. Most of the double-stranded segments are formed from transcription of DNA by intramolecular base-pairing of inverted complementary sequences separated by a single-stranded loop. Some double-stranded segments of RNA are normal in all organisms.	Double-Stranded RNA,Double Stranded RNA,RNA, Double Stranded
D012367	RNA, Viral	Ribonucleic acid that makes up the genetic material of viruses.	Viral RNA
D012401	Rotavirus	A genus of REOVIRIDAE, causing acute gastroenteritis in BIRDS and MAMMALS, including humans. Transmission is horizontal and by environmental contamination. Seven species (Rotaviruses A thru G) are recognized.	Neonatal Calf Diarrhea Virus,Rotaviruses