A panel of outstanding investigators surveys and explains the major cutting-edge methods used in thryroid receptor (TR) research and explains their practical experimental details. Described in step-by-step detail to ensure robust experimental results, the techniques presented cover a wide variety of key areas, including TR in development and knockout (mouse and Xenopus), transcriptional regulation by TRs in both cell-free systems and in living cells, and TR mutant analysis of patients. Additional methods provide powerful tools for the isolation of TR-regulated protein complexes, for studying...

There have been many important advances in our understanding of biological membrane structure and function over the last decade. Much of this progress has been driven by the development of new techniques for studying membrane components and their interactions. Tradition- ally, the investigation of membranes has largely occurred within the domains of biochemistry, physical chemistry, and cell biology; but many of the most significant advances have resulted from an expansion into other disciplines, such as molecular biology, immunology, and the clinical sciences. In these two volumes we have...

The natural, biological, medical, and related sciences would not be what they are today without the microscope. After the introduction of the optical microscope, a second breakthrough in morphostructural surface analysis occurred in the 1940s with the development of the scanning electron microscope (SEM), which, instead of light (i. e., photons) and glass lenses, uses electrons and electromagnetic lenses (magnetic coils). Optical and scanning (or transmission) electron microscopes are called "far-field microscopes" because of the long distance between the sample and the point at which the...

The past decade has seen an extraordinary growth in research interest in neurotrophic factors, and the study of the neurotrophin family has led this activity. Nevertheless, this area of research has often struggled as a result of techniques that were either inadequate or just emerging from other research fields and disciplines. Neurotrophin Protocols has brought together many leaders in the neurotrophin field who detail their special expertise in a wide variety of techniques. Though most procedures are valid across many diff- ent fields of research, some of those described here have been...

This is the second of three volumes of Methods in Molecular Biology that deal with Physical Methods of Analysis. The first of these, Spectroscopic Methods and Analyses dealt with NMR spec- troscopy, mass spectrometry, and metalloprotein techniques, and the third will cover X-ray crystallographic methods. As with the first volume. Microscopy, Optical Spectroscopy, and Macroscopic Techniques is intended to provide a basic understand- ing for the biochemist or biologist who needs to collaborate with spe- cialists in applying the techniques of modern physical chemistry to biological...

0ne 0fthe many 1mpact5 0frec0m61nant DNA techn0109y 0ver the 1a5t 15 year5 ha5 6een a 5tr0n91y refre5hed 1ntere5t 1n meth0d5 f0r the 5eparat10n and pur1f1cat10n 0f pr0te1n5.7h15 1ntere5t ha5 enc- pa55ed n0t 0n1y ana1yt1ca1 5eparat10n5, 6ut a150 5ma11- and 1ar9e-5ca1e preparat1ve meth0d5 d1rected t0 60th pure and app11ed re5earch thr0u9h0ut 610109y and 610med1c1ne. Many 0f the new 0r 5u65tant1a11y m0d1f1ed techn14ue5 deve1- 0ped have 6een rep0rted 1n the 11terature, 6ut a 5uff1c1ency 0f deta11ed pract1ca1 he1p 1n e5ta6115h1n9 the5e meth0d5 f0r the f1r5t t1me 1n a new 1a60rat0ry ha5 0ften 6een...

Advances in bioscience research usually arise as a result of the continu- ing refinement of existing technologies. However, there are a number of occa- sions v DEGREESrhere newly developed methodologies have a profound effect on nearly all areas of research. Frequently these are techniques that are elegantly simple in concept and require minimal technical manipulation. Two of these revolu- tionary techniques are the focus ofPCR Sequencing Protocols. The first such technique is enzymatic chain termination sequencing developed by Sanger and his co-workers in Cambridge and reported in 1977. This...

The first libraries of complementary DNA (cDNA) clones were con- structed in the mid-to-late 1970s using RNA-dependent DNA polymerase (reverse transcriptase) to convert poly A* mRNA into double-stranded cDNA suitable for insertion into prokaryotic vectors. Since then cDNA technology has become a fundamental tool for the molecular biologist and at the same time some very significant advances have occurred in the methods for con- structing and screening cDNA libraries. It is not the aim of cDNA Library Protocols to give a comprehensive review of all cDNA library-based methodologies; instead we...

The development of recombinant DNA technology and methods for transferring recombinant genes into plants has brought about significant advances in plant science. First, it has allowed investigation, using reporter genes, into the transcriptional regulation of plant genes-a key to the under- standing of the biochemical basis of growth and development in plants. Second, gene transfer technology has facilitated the molecular cloning, by tagging genomic sequences, of important genes (e. g., homeotic genes) whose gene products control the normal pattern of growth and differentia- tion of plants....

The ability to introduce macromolecules into animal cells, includ- ing DNA, RNA, proteins, and other bioactive compounds has facili- tated a broad range of biological studies, from biochemistry and biophysics to molecular biology, cell biology, and whole animal stud- ies. Gene transfer technology in particular will continue to play an essential role in studies aimed at improving our understanding of the relationships between the gene structure and function, and it has impor- tant practical applications in both biotechnology and biomedicine, as evidenced by the current intense interest in gene...