A collection of standard and cutting-edge techniques for using Xenopus oocytes and oocytes/egg extracts to reconstitute biological and cellular processes. These readily reproducible methods take advantage of the oocyte's impressive protein abundance, its striking protein translation capacity, and its breathtaking possibilities for the assembly of infectious viral particles by single cell injection of multiple RNAs. The authors focus on the versatility of frog oocytes and egg extracts in cell biology and signal transduction, and cover all the major uses of oocytes/extracts as experimental models.
1. Resources for genetic and genomic studies of Xenopus; Steven Klein; 2. The physiology of the Xenopus ovaries; Stephen Hammes; 3. Oocyte isolation and enucleation; Shawn Xun Liu; 4. Xenopus tropicalis oocytes: more than just a beautiful genome; Nick Duesbery; 5. Oocyte expression with injection of purified T7 RNA polymerase; Michel DeWaard; 6. Visualization of the cytoskeleton in Xenopus oocytes and eggs by confocal immunofluorescence microscopy; David Gard; 7. Multiphoton laser scanning microscopy as a tool for Xenopus oocyte Research; James Lechleiter; 8. Imaging Ca2+ signals in Xenopus oocytes; Ian Parker; 9. Chromosomal DNA replication in a soluble cell-free system derived from Xenopus eggs; Johannes Walter; 10. Chromatin assembly of DNA templates micro-injected into Xenopus oocytes; Genevieve Almouzni; 11. Pre-mRNA splicing in the nuclei of Xenopus oocytes; Yi-Tao Yao; 12. Chromatin immunoprecipitation for studying transcriptional regulation in Xenopus oocytes and tadples; Jiemin Wong; 13. Cytoplasmic mRNA polyadenylation and translation assays; Raul Mendez; 14. Xenopus egg extracts - a model system to study proprotein convertases; Kathleen Shannon; 15. Analysis of molecular chaperones using a Xenopus oocyte protein refolding assay; John Heikkila; 16. Ubiquitin-mediated protein degradation in egg extracts; Thierry Lorca; 17. Introduction to nucleo-cytoplasmic transport; Reiner Peters; 18. Use of Xenopus oocyte nuclei and nuclear envelopes in nucleocytoplasmic transport studies; Reiner Peters; 19. Nuclear pore complex (NPC) structure and plasticity revealed by electron microscopy and atomic force microscopy; Ueli Aebi; 20. In vitro study of nuclear assembly and nuclear input using Xenopus egg. extracts; Douglas Forbes; 21. Use of intact Xenopus oocytes in nucleocytoplasmic transport studies; Nelly Pante; 22. Studying the mechanosensitivity of voltage-gated channels using oocyte patches; CE Morris; 23. Oocytes as an expression system for studying drug and pesticide receptor/channel targets; David Sattelle; 24. Microtransplantation of membranes for neurotransmitter receptor over-expressing cells to Xenopus oocytes; Fabrizio Eusebi and Richardo Miledi; 25. Reconstitution of Golgi disassembly by mitotic Xenopus egg extracts in semi-intact MDCK cells; Fumi Kano; 26. Exploring RNA virus assembly in Xenopus oocytes; Raul Andino; 27. Studying apoptosis in vitro using the Xenopus egg extract reconstitution system; Sally Kornbluth; 28. Signal transduction of egg fertilization: focused proteomics on membrane/lipid rafts; Ken-ichi Sato; 29. Localized sampling, electrophoresis and biosensor analysis of Xenopus laevis cytoplasm for subcellular biochemical assays); Christopher E. Sims; 30. Measuring PKA activities in live oocytes; Jing Wang; 31. Using Xenopus oocyte extracts to study signal transduction; Joan Ruderman; 32. Oocyte extracts for the study of meiotic M-M transition; Keita Ohsumi; 33. Studying spindle dynamics in Xenopus egg extracts; Rebacca Heald.
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