Plant Cell Electroporation And Electrofusion Protocols

1. Front Matter

   Pages i-xiv

   PDF

2. Theory and Instrumentation

   1. Electroporation Theory

      • James C. Weaver

      Pages 3-28

   2. Effects of Pulse Length and Strength on Electroporation Efficiency

      • Sek Wen Hui

      Pages 29-40

   3. Instrumentation

      • Gunter A. Hofmann

      Pages 41-59

3. Electroporation Protocols

   1. Electroporation of Agrobacterium tumefaciens

      • Amke den Dulk-Ras, Paul J. J. Hooykaas

      Pages 63-72

   2. Electroporation of DNA into the Unicellular Green Alga Chlamydomonas reinhardtii

      • Laura R. Keller

      Pages 73-79

   3. Pollen Electrotransformation in Tobacco

      • James A. Saunders, Benjamin F. Matthews

      Pages 81-88

   4. Electroporation of Tobacco Leaf Protoplasts Using Plasmid DNA or Total Genomic DNA

      • Patrick Gallois, Keith Lindsey, Renee Malone

      Pages 89-107

   5. Electroporation of Brassica

      • Frank Siegemund, Klaus Eimert

      Pages 109-120

   6. Transformation of Maize by Electroporation of Embryos

      • Carol A. Rhodes, Kathleen A. Marrs, Lynn E. Murry

      Pages 121-131

   7. Transient Gene Expression Analysis in Electroporated Maize Protoplasts

      • Kathleen A. Marrs, J. C. Carle Urioste

      Pages 133-145

   8. Reporter Genes and Transient Assays for Plants

      • Benjamin F. Matthews, James A. Saunders, Joan S. Gebhardt, Jhy-Jhu Lin, Susan M. Koehler

      Pages 147-162

4. Electrofusion Protocols

   1. Electrofusion of Plant Protoplasts

      • Harold N. Trick, George W. Bates

      Pages 165-179

   2. Protoplast Electrofusion and Regeneration in Potato

      • Jianping Cheng, James A. Saunders

      Pages 181-188

   3. Polymer-Supported Electrofusion of Protoplasts

      • Lei Zhang

      Pages 189-202

5. Back Matter

   Pages 203-205

   PDF

Gene transfer is an essential technology for improving our under­ standing of gene structure and function. Although there are many meth­ ods by which DNA may be introduced into cells—including heat and chemical treatments, and microinjection—electroporation has been found to be the most versatile gene transfer technique. Electroporation is effective with a wide variety of cell types, including those that are difficult to transform by other means. For many cell types, electroporation is either the most efficient or the only means known to effect gene transfer. The early and broad success of electric field-medi­ ated DNA transfer soon prompted researchers to investigate electroporation for transferring other types of molecules into cells, in­ cluding RNA, enzymes, antibodies, and analytic dyes. The first section of Plant Cell Electroporation and Electrofusion Protocols includes two chapters that serve as a guide to theoretical and practical aspects of electroporation, and will be of particular interest to those developing protocols for as yet untested species or cell types, and a third chapter that describes commercially available electroporation instruments. The remaining chapters describe well-tested protocols for DNA electrotransfection, electroporation of other biomolecules, or cell electrofusion. These chapters also include brief discussions of alterna­ tives to electric field-based methods, citing the advantages and limita­ tions of the various methods for achieving specific goals.

• Harvard University, Boston

  Jac A. Nickoloff

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