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A Safety Considerations Many techniques described here involve a number of hazards, such as high electrical current and voltage, radioactivity and highly toxic chemicals. It is absolutely essential that the instructions of equipment manufacturers be followed, and that particular attention be paid to the local and federal safety regulations. B Introduction The expression of prokaryotic and eukaryotic genes has been shown most often to be regulated at the level of mRNA synthesis. Thanks to the rapid development of methods for dissecting DNA sequences, cis-acting regulatory elements such as promoters and enhancers have been recognised. More recently, the widely expressed intuition that discrete sequences within these elements constitute binding sites for sequence-specific binding proteins has been confirmed, especially through the use of "footprinting" assays (for examples, Galas and Schmitz, 1978). This and similar assays have already resulted in the recognition, isolation and analysis of DNA-bind ing proteins for several genes. Excellent reviews exist of the structural studies on these transcription regulatory proteins and related DNA elements (for example, Glover, 1989 and Johnson and McKnight, 1989), to which the reader is referred for detailed information. To set the scene for applications of the techniques described in this volume, only the barest outline of previous studies is presented here. Protein-DNA interactions are dependent on very specific tertiary configurations of the binding protein which allow the closest contact with the DNA helix.
First multi-year cumulation covers six years: 1965-70.
The occurrence of 5-methylcytosine in DNA was first described in 1948 by Hotchkiss (see first chapter). Recognition of its possible physiologi cal role in eucaryotes was first suggested in 1964 by Srinivasan and Borek (see first chapter). Since then work in a great many laboratories has established both the ubiquity of 5-methylcytosine and the catholicity of its possible regulatory function. The explosive increase in the number of publications dealing with DNA methylation attests to its importance and makes it impossible to write a comprehensive coverage of the literature within the scope of a general review. Since the publication of the 3 most recent books dealing with the subject (DNA methylation by Razin A. , Cedar H. and Riggs A. D. , 1984 Springer Verlag; Molecular Biology of DNA methylation by Adams R. L. P. and Burdon R. H. , 1985 Springer Verlag; Nucleic Acids Methylation, UCLA Symposium suppl. 128, 1989) considerable progress both in the techniques and results has been made in the field of DNA methylation. Thus we asked several authors to write chapters dealing with aspects of DNA methyla tion in which they are experts. This book should be most useful for students, teachers as well as researchers in the field of differentiation and gene regulation. We are most grateful to all our colleagues who were willing to spend much time and effort on the publication of this book. We also want to express our gratitude to Yan Chim Jost for her help in preparing this book.
Although DNA fingerprinting is a very young branch of molecular genetics, being barely six years old, its recent impact on science, law and politics has been dramatic. The application of DNA finger printing to forensic and legal medicine has guaranteed a high public profile for this technology, and indeed, scarcely a week goes by with out the press reporting yet another crime successfully solved by molec ular genetics. Less spectacularly, but equally importantly, DNA typing methods are steadily diffusing into an ever wider set of applications and research fields, ranging from medicine through to conservation biology. To date, two DNA fingerprinting workshops have been held in the UK, one in 1988 organised by Terry Burke at the University of Leicester, and the second in 1989 at the University of Nottingham, co-ordinated by David Parkin. In parallel with these workshops, which have provided an important focus for researchers, Bill Amos and Josephine Pemberton in Cambridge have established an informal newsletter "Fingerprint News" which is playing a major role as a forum for DNA fingerprinters. By 1989, it was clear that the field had broadened sufficiently to warrant a full international meeting. As a result, Gaudenz Dolf took on the task of organising the first, of what I hope will be many, International Symposium of DNA Fingerprinting held at Bern during Ist-3rd October 1990. The success of the meeting can be judged from the remarkable attendance, with 270 delegates from no less than 30 countries.
Cell Biology: A Laboratory Handbook, Volume 3 is a handbook on cell biology and covers topics ranging from transfer of macromolecules and small molecules to cloning of embryos, transgenics, and gene targeting. Cell-free extracts, permeabilized cell systems, and expression systems are also discussed, along with proteins. Comprised of 58 chapters, this volume begins with a detailed account of microinjection of RNA, DNA, and proteins into somatic cells, followed by an analysis of computer-automated capillary microinjection of macromolecules into living cells. The reader is then introduced to syringe loading as a method for inserting macromolecules into cells in suspension; electroporation of cells; and the use of liposomes in drug targeting. Subsequent chapters focus on the cloning of rabbit embryos by nuclear transplantation; gene targeting by homologous recombination in embryonic stem cells; production and isolation of recombinant viruses; and gel electrophoresis. This book will be of interest to geneticists and molecular biologists.
Since the appearance of photosynthesis on our planet, all living organisms have been facing a new abundant, extremely reactive element, oxygen. This element is used for the synthesis of highly energetic compounds, but can also generate molecules capable of damaging constituents of living structures, including proteins, nucleic acids and lipids. It is therefore no surprise that all biological organisms have evolved multiple and sophisticated ways to reduce the detrimental effects of oxygen. For cells and tissues of highly organized animals, particularly cells of the immune system, superoxidation products play an important role, via oxidative stress, in activation, inflammation and viral and bacterial infections. We must be grateful to Drs. C. Pasquier, C. Auclair, L. Packer and R. Olivier for having brought together many specialists in an international meeting held in Paris in March 1993, at the Ministry of Research. This book comprises an impressive amount of recent knowledge, a real mine for the reader in this fast developing field of research. Furthermore, we should not forget that this basic research may lead in the future to new therapeutic approaches to the most important pathologies of the latter part of this century, including AIDS.