附註:Includes bibliographical references and index.
Intro; Contents; List of protocols; Abbreviations; 1 Investigation of RNA virus genome structure; 1 Introduction-the nature of the virus genome; Properties of the genomes of RNA viruses; Properties of the virions of RNA viruses; Paradigm for analysis of RNA virus genome structure; 2 Growth, assay, and purification of RNA viruses; Source of virus: in vivo versus in vitro methods; Assay of virus yield; Harvesting and concentrating of virus; Purification of virus; Radiolabelling; 3 RNA extraction; Genome RNA extraction from virions; Genome RNA extraction without virion purification
4 Fractionation of RNA, and analysis by Northern (RNA) blottingFractionation of RNA; Northern blotting and detection with labelled probes; 5 Further analysis; Characterization by restriction-endonuclease digest patterns; Characterization by ribonuclease protection; References; 2 Mutagenesis of RNA virus genomes; 1 Introduction; 2 Generation of RNA virus mutants; Selection of mutants from the RNA quasispecies; Recombination of RNA virus genomes; Reassortment of segmented RNA virus genomes; Generation of defective RNA virus populations by passage in culture
Recovery of RNA virus mutants from infectious cDNA3 Determining the genotype of RNA virus mutants; 4 Analysis of the frequency of RNA virus mutants; References; 3 Analysis of transcriptional control In RNA virus Infections; 1 Introduction; 2 Analysis of virus mRNAs in virus-infected cells; General considerations; Radiolabelling and extraction of virus RNAs; Gel electrophoresis of RNAs; 3 Analysis of virus mRNA structure; General considerations; Separation of virus RNA in preparative agarose gels; Extraction of virus RNA from gel slices; One-dimensional oligonucleotide fingerprinting
Northern blot hybridization4 The use of RNA reporter constructs for transcriptional assays; RNA transcription in vitro; RNA transfection of DI RNA construct and helper virus infection; DNA transfection and vaccinia virus infection; CAT reporter assay system; Detection and quantitation of minute amount of virus RNAs; References; 4 Analysis of RNA virus-encoded proteinases; 1 Introduction; 2 Defining the proteinase type; Inhibitor studies; Sequence analysis; 3 Delimiting the proteinase (domain); Deletion/truncation analysis; Screening for proteolytic activity using translation systems in vitro
4 Bacterial expressionExpression of inactive proteinase; Affinity 'tagging'; Insolubility of expressed proteinases; 5 Substrate specificity; 6 Artificial 'reporter' polyproteins; References; 5 Detection and analysis of host gene targets for oncogenic retroviruses; 1 Introduction; 2 Insertional mutagenesis; Analysis of virus integration patterns; Cloning of virus integration sites; Analysis of host DNA flanking the provirus integration sites; Location of genes at integration sites; 3 Transduction of virus oncogenes; Properties of transducing viruses; Biological methods of detection
摘要:RNA Viruses: A Practical Approach provides a broad treatment of the principles and practice of RNA virus research to ensure the widest possible audience. RNA Viruses: A Practical Approach will be of interest to all laboratories involved in virus culture, including both academic and industrial research groups and public health laboratories.
