Gene Transfer and Expression in Mammalian CellsS.C. Makrides Newnes, 24 okt 2003 - 722 pagina's The production of proteins in mammalian cells is an important tool in numerous scientific and commercial areas. For example, proteins for human therapy, vaccination or diagnostic applications are typically produced in mammalian cells. Gene cloning, protein engineering, biochemical and biophysical characterization of proteins also require the use of gene expression in mammalian cells. Other applications in widespread use involve screening of libraries of chemical compounds in drug discovery, and the development of cell-based biosensors.This book presents a state-of-the-art comprehensive coverage of the technical aspects of gene expression in mammalian cells, written by experienced scientists working at the forefront of the field. |
Inhoudsopgave
Expression in primary cell cultures | |
Expression in hippocampal slice cultures | |
SFV vectors in vivo | |
Safety of SFV vectors | |
SFV vectors in gene therapy | |
SFV vectors as tools for virus assembly | |
Introduction 2 Baculoviruses as insect cell expression vectors 3 Molecular biology of virus vector construction | |
Novel technologies | |
Gene delivery | |
Virusbased vectors for gene expression in mammalian | |
Gene expression | |
Production and analyses of viral particles | |
Conclusion | |
Vaccinia virus | |
Introduction | |
Vaccinia virus molecular biology | |
Construction of recombinant poxvirus vectors | |
Chimeric VVbacteriophage expression vectors | |
Improved safety of VV vectors | |
Nonvaccinia poxvirus vectors | |
Laboratory and clinical applications | |
Baculovirus | |
Baculoviruses as mammalian cell expression vectors | |
Characteristics of baculovirusmediated mammalian gene delivery | |
Applications of the baculovirus mammalian gene delivery vector | |
Points to consider | |
Assay development | |
Coronavirus | |
Introduction | |
Coronavirus pathogenicity | |
Molecular biology of coronavirus | |
Helperdependent expression systems | |
Single genome coronavirus vectors | |
Optimization of transcription levels | |
Modification of coronavirus tropism and virulence | |
Expression systems based on arteriviruses | |
Conclusions | |
Poliovirus | |
Sindbis virus 1 Introduction | |
Viral genetic elements | |
Packaging of replicons | |
Effects of alphaviruses and replicons on infected cells | |
Expression in neurons | |
Insect and crustacea | |
Vaccines for infectious diseases and cancer | |
Perspectives | |
Semliki Forest virus | |
Introduction | |
Expression of topologically different proteins | |
Host cell range | |
Scaleup of protein production | |
Conclusions | |
Retrovirus | |
Introduction | |
Biology of retroviruses | |
Development of recombinant retrovirus vectors | |
A poliovirus vectorbased HIV vaccine 3 Poliovirus vector development and its immunogenic potential | |
Expression of the transgene | |
Targeting | |
Conclusion and perspectives | |
Lentiviruses | |
Introduction | |
Genetic structure and biology of lentiviruses | |
HIV1derived vector packaging system | |
Lentiviral vector production | |
Pseudotyped vectors | |
Expression from lentiviral vectors | |
Gene transfer applications 8 Conclusion | |
Methods for DNA introduction into mammalian cells | |
Barriers to successful transfection | |
Comparison of available methods | |
Applications | |
Lipid reagents for DNA transfer into mammalian cells | |
Introduction | |
General structure of cationic lipids | |
Formulation and physicochemical properties | |
Mechanisms of in vitro transfection | |
In vivo administration | |
Targeting | |
New strategies to improve in vivo transfection | |
Conclusion | |
Reporter genes for monitoring gene expression in mammalian cells | |
Introduction | |
Reporter gene vectors and fusions | |
Green fluorescent protein | |
Luciferases | |
Alkaline phosphatase | |
Chloramphenicol acetyltransferase | |
βGalactosidase | |
Gene transfer and gene amplification in mammalian cells | |
Cotransfer of multiple plasmidsviruses as an attractive | |
Use of scaffoldmatrixattachment regions for protein | |
Chromatin insulators and position effects | |
Locus control regions | |
Pathways and functions of mammalian protein | |
Metabolic engineering of mammalian cells for higher | |
Optimization of plasmid backbone for gene expression | |
Protein production in transgenic animals | |
Strategies for the purification of recombinant proteins | |
Product recovery and initial purification | |
Purification of proteins from culture media | |
Purification of proteins from cell lysates | |
Volume reduction and partial purification | |
Chromatographic purification of recombinant proteins | |
Product stability and longterm storage | |
Subject Index | |
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Veelvoorkomende woorden en zinsdelen
AAV vectors Acad adenovirus animals antibody antigens applications baculovirus binding Biol Biotechnol capsid capsid proteins cDNA cell lines cell types cellular cloning containing coronavirus culture cytotoxic deleted demonstrated disease EBV-derived efficient encoding expression cassette expression levels expression systems expression vectors function gene delivery gene expression Gene Ther gene therapy gene transfer genetic genome glycoprotein helper virus host cell human immune response induce infected cells inserted J.C. Glorioso latent lytic mammalian cells membrane mice minigenomes monkeys mRNA mucosal mutant Natl neurons oriP packaging pathogenic plasmid poliovirus Proc production promoter receptor recombinant proteins replication replication-competent replicon reporter gene rSV40 vectors sequences SFV vectors Sindbis virus stable studies synthesis target TGEV tissue titers transcription transduction transfection transgene transgene expression tumor vaccine vaccine vectors viral DNA viral genes virion Virol Virology viruses vitro vivo wild-type