Yakshe Krystle A, Franklin Zachary D, Ball Judith M
Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, College Station, Texas.
Curr Protoc Microbiol. 2015 May 1;37:15C.6.1-44. doi: 10.1002/9780471729259.mc15c06s37.
Rotavirus (RV) contains 11 double-stranded RNA segments that encode for twelve structural and nonstructural proteins. The separation and isolation of viral RNA is a necessary precursor for many experimental techniques and can be useful for rapid RV RNA typing and sequencing of different rotavirus strains. The segmented genome enables RV to recombine easily. These recombinant viruses are essential for many purposes, including generation of potential vaccine strains. Rotavirus gene 10 expresses the viral enterotoxin, NSP4, which has been the focus of several studies due to the influence of NSP4 on rotavirus replication, morphogenesis, and pathogenesis. This unit will describe the isolation and separation of viral RNAs, the production characterization of recombinant RV in culture, and the expression and isolation of NSP4 in mammalian and insect cells.
轮状病毒（RV）含有11个双链RNA片段，编码12种结构蛋白和非结构蛋白。病毒RNA的分离和纯化是许多实验技术的必要前提，并且有助于对不同轮状病毒株进行快速的RV RNA分型和测序。分段基因组使RV易于重组。这些重组病毒在许多方面都至关重要，包括潜在疫苗株的产生。轮状病毒基因10表达病毒肠毒素NSP4，由于NSP4对轮状病毒复制、形态发生和发病机制的影响，它一直是多项研究的重点。本单元将描述病毒RNA的分离和纯化、培养中重组RV的生产特性，以及NSP4在哺乳动物细胞和昆虫细胞中的表达与分离。
