Showing papers in "Methods in 1998"

TL;DR: Precise DNA rearrangements and genetic switches can be efficiently generated in a straightforward manner using Cre recombinase and are likely to have a profound impact on developmental biology and the generation of useful animal models of human disease.

TL;DR: This article describes, in detail, how to generate and characterize GAL4 lines and how to prepare UAS-target gene lines, and considers the range of U AS-reporters currently available and several new modifications of the GAL3 system.

TL;DR: Six procedures are given for preservation of myelinated nerve fibers for light or electron microscopic studies, and axon/myelin coherence is best preserved by decreasing the temperature during the rinse with saline.

TL;DR: Simulation of the diffusional encounter of the extracellular ribonuclease, barnase, and its intracellular inhibitor, barstar, shows how experimental rates for a series of mutants and the dependence of rates on ionic strength can be reproduced well by Brownian dynamics simulations.

TL;DR: Strains carrying these genetically marked FLP/FRT chromosomes have greatly enhanced the ability to study gene function in both germline and somatic Drosophila cells.

TL;DR: All the steps required to produce a virus carrying a transgene of interest and the methodologies behind designing and carrying out misexpression strategies are outlined, and some useful techniques for analyzing infected embryos are described.

TL;DR: A novel one-step procedure for large-scale preparation of individual histones and their acetylated isoforms for the analysis of substrate and site specificity of the enzymes is presented.

TL;DR: It is emphasized that some of the conclusions about neuronal-astrocytic interactions reached on the basis of studies in cultured cells and confirmed in intact tissue may not yet have been completely integrated into general neuroscience knowledge.

TL;DR: The mechanisms by which Tax regulates viral and cellular gene expression are discussed, including modulation of gene expression via the ATF/CREB and NF-kappaB pathways is linked to its transforming properties.

TL;DR: Improved knowledge of stacking interactions may lead to a better understanding of the architecture and inherent flexibility of particular DNA sequences and may provide insight into the principles that dictate the structural changes and specificity patterns observed in the binding of some intercalating ligands to DNA.

TL;DR: The identification and characterization of four native nuclear high-molecular-weight HAT complexes in Saccharomyces cerevisiae can be used in a variety of functional assays to further address questions of how acetylation has an impact on transcriptional regulation.

TL;DR: Methods and strategies used in the authors' laboratories to study phosphorylation and activation of PDE3B in adipocytes and in vitro are described and presumably allowing close coupling of the regulation of steady-state concentrations of both cAMP and PKA and, thereby, control of lipolysis.

TL;DR: The use of replication-incompetent retroviral vectors for the analysis of lineal relationships in developing vertebrate tissues and the strategies and current methods in use in the laboratory are described.

TL;DR: These cultures are most commonly derived from perinatal rat brain, but a protocol for preparation from mouse brain is also provided because of the increasing number of studies that use mice to facilitate molecular biological techniques.

TL;DR: Comparative modeling, or modeling by homology, is still the method of choice when the unknown protein shares any significant sequence similarity with a protein of known structure.

TL;DR: Methodology for the isolation and maintenance of microglia in cell cultures prepared from murine and feline animals is described and a strategy is presented that proposes to use the microglial cell lineage to effectively deliver therapeutic compounds to the CNS from the peripheral circulation.

TL;DR: The cells in vitro and following transplantation into the neonatal SVZa and adult striatum are found to continue both to proliferate and to differentiate into neurons, suggesting that the unique features of the SVZA progeny are specified intrinsically rather than by their extrinsic environment.

TL;DR: While investigating the localization of synaptophysin in PC12 cells using immunofluorescence microscopy, a striking difference in its apparent subcellular distribution depending on whether digitonin or Triton X-100 was used as permeabilization agent of paraformaldehyde (PFA)-fixed cells was found.

TL;DR: This work describes first the resolution of different PDEs using high-performance anion-exchange chromatography and then a Western blotting methodology for identifying or authenticating PDE1 activities, and presents an immunoprecipitation method that can be used for quantitating specific Pde1 isoforms.

TL;DR: A cell-free reconstitution system was developed that generates vesicles from PC12 membrane precursors in the presence of ATP and brain cytosol and is temperature dependent, and vesiculation and sorting can be studied in this cell- free system.

TL;DR: Results indicate the importance of a specific moncyte stimulus in the generation of a unique HCMV permissive macrophage phenotype as well as why virus is commonly reactivated in transplant patients.

TL;DR: Those approaches to chemical toxicity that have taken a more "personalized" configuration and have undergone implementation into software programs able to perform the various steps of the assessment of the hazard posed by the chemicals are discussed.

TL;DR: A baculovirus-insect cell system is developed for high-yield expression of the small and medium isozymes and their mutants using the insect cell system and methods for studying 2-5(A) synthetase-dsRNA interactions and protein-protein interactions among the subunits of the two isoz enzymes are reported.

TL;DR: The use of much larger DNA fragments cloned in yeast artificial clones (YACs), bacterial artificial clones, or P1-based artificial clones has been found to give much better levels of expression, generally very close to that of an endogenous gene, and tissue-specific expression matching that of the endogenous gene.

TL;DR: Several cell-free systems are described for studies of regulated exocytosis derived from PC12 cells, clonal cell lines of adrenal medullary origin, that possess large dense-core vesicles that retain their competence for regulated excytosis in a variety of permeable cell and isolated membrane preparations.

TL;DR: The iterative approach employed to describe alpha 1b-AR activation in terms of molecular structure and dynamics allows further complications of the model to allow prediction and interpretation of an ever-increasing number of experimental data.

TL;DR: The results show that embryonic cortical cells exhibit a range of naturally occurring buoyant densities that can be used to expeditiously fractionate cortical cells according to their pre- or postmitotic status, thus providing ready access for cellular and molecular studies of proliferation and differentiation.

TL;DR: Using an in vitro cosedimentation assay, it is demonstrated that MxA tightly interacts with components of the ribonucleoprotein complex of Thogoto virus, an influenza-like virus transmitted by ticks.

TL;DR: A number of experimental approaches that have recently been developed for studying the interactions between catalytic and inhibitory subunits of PDE6, as well as the dynamics of cGMP binding to and dissociation from the PDE 6 noncatalytic sites are described.

TL;DR: A standard method for microwave-stimulated antigen retrieval in formaldehyde-fixed paraffin-embedded and nonembedded tissue is presented that results, in general, in very good staining for antibodies used in neuroscience.