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Molecular Plant Biology is an all-new replacement for the original Practical Approach book Plant Molecular Biology that was first published in 1988. The rapid advances made in plant sciences during the past decade are reflected by the need to produce a two-volume book to cover all the relevant methodologies. The new book incorporates many of the fundamental procedures outlined in the original book, but these are fully updated to reflect advances technology and the development of new methodologies. The new books also incorporate many approaches that were not available in the earlier volume. Each chapter has been written by an international expert with current practical expertise in the topics covered. Each book is divided into three sections. The approaches outlined in Volume One cover a wide range of techniques for gene isolation, gene identification and subsequent isolation, as well as for studies of gene organization. The methods described range from classical mutagenesis through plant transformation, T-DNA and transposon tagging strategies, genomic subtraction, gene mapping, construction and screening of YAC, BAC and cosmid libraries chromosome in situ, and isolation of cDNA sequences by western and southwestern library screens, and complementation cloning. Volume Two focuses on experimental approaches for studies on gene expression, gene product analysis, with the final section devoted to emerging technologies. Topics covered include a range of techniques for transcript analysis, including In situ Hybridization and DNA microarrays. DNA-protein interaction methods are also covered in detail. Inducible gene expression in plants as well as expression and analysis of recombinant proteins, and analysis of protein import into chloroplasts are covered as well as techniques for fractionation of plant tissue for biochemical analyses and the study of protein-protein interactions with the yeast two-hybrid system. A range of approaches for using antibodies as tools are also described including the use of antibody phage display libraries. The final section on emerging technologies describes methodologies for calcium imaging and for the spatial and temporal analysis of reporter genes such as luciferase and green fluorescent protein. The final area covers a range of experimental procedures for moss, which is emerging as a new model organism.
Part 1 Gene Expression: 1 Transcript analysis; 2 In situ hybridization; 3 DNA microarrays for gene expression analysis; 4 DNA-protein interactions; 5 Inducible gene expression in plants. Part 2 Gene Product Analysis: 6 Expression of recombinant proteins; 7 Import of proteins into isolated chloroplasts and thylakoid membranes; 8 Fractionation of plant tissue for biochemical analysis; 9 Studying protein-protein interactions with the yeast two-hybrid system; 10 Antibody techniques; 11 Antibody phage display libraries. Part 3 Emerging Technologies: 12 Spatial and temporal measurements of calcium ions in living plant cells; 13 Reporter genes for plant cells; 14 Moss gene technology.
The chapters are well written, with sufficient background detail and a step-by-step guide to each technique. Overall, this will be a useful manual for any plant molecular biology laboratory. Microbiology Today