Showing papers in "Methods in 1990"

TL;DR: The streak seeding technique as mentioned in this paper can be used to bypass the nucleation step and thus accomplish the decoupling between nucleation and crystal growth in order to grow large, regular crystals.

TL;DR: In this paper, the authors discuss the possibility that the T7 RNAP results indicate that glycerol and polyhydric alcohols may be generally useful in crystallizing proteins in which conformational flexibility is a problem.

TL;DR: PFGE and blot hybridization analyses of Arabidopsis DNA-containing yeast artificial chromosomes demonstrated that both unique and highly repeated DNA sequences are represented.

TL;DR: This article addresses how the physical behavior of the detergent, in the form of micelles or protein-detergent aggregates, affects crystallization and concludes that many integral membrane proteins could be crystallized if pure and monodisperse preparations in a suitable detergent system can be prepared.

TL;DR: A set of conditions for initial cocrystallization trials with a new DNA binding protein is suggested, and detailed protocols for the purification of chemically synthesized fragments are suggested.

TL;DR: In this paper, two-level factorial, fractional factorial and generalized multilevel incomplete factorial (GMLF) designs for screening crystal growth conditions are compared and evaluated.

TL;DR: In this paper, a dual-plasmid expression system that co-expresses Saccharomyces cerevisiae myristoyl-CoA:protein NMT and various known mammalian NMT proteins in Escherichia coli is described.

TL;DR: In this article, the translational diffusion coefficient (D) of a protein is estimated using dynamic light scattering (DLS) and the protein concentration dependence of D is correlated with the thermodynamic equilibrium governing these two types of aggregates.

TL;DR: The purification procedure is reproducible and enables the isolation of microgram amounts of purified enzyme from 50 rat brains, and two methods for assaying enzymatic activity are described.

TL;DR: The purpose of this article is to present a brief description of protein acylation and prenylation modifications that have been observed and the details of how to detect the modifications by metabolic radiolabeling of cellular proteins.

TL;DR: Analytical methods to determine the presence of a prenyl modification, the structure of the preNyl group, and the lipid: protein molar ratio for candidate proteins or their proteolytic fragments are described.

TL;DR: Methods for the characterization of glycosylinositol phospholipid anchors of membrane proteins isolated from metabolically labeled cells, followed by the separate analysis of the glycan and lipid components are described.

TL;DR: In this article, a detailed view of the crystallization process and several representative examples of the crystal crystallization of oligonucleotides are presented. But the success depends critically on the probability of obtaining suitable crystals, which appears to be not very high.

TL;DR: In this article, it was shown that solubility is affected more by anions than by cations and that the effectiveness of the anions is in an order roughly the reverse of that of the Hofmeister series.

TL;DR: The synthesis and characterization of AFC are reported and it is demonstrated how it has exploited its distinctive properties to develop a rapid assay for carboxyl methyltransferase activity.

TL;DR: The Biological Macromolecule Crystallization Database (BMCD) as mentioned in this paper contains the crystallization conditions and crystal data for crystal forms of all classes of biological macromolecules for which the unit cell dimensions and space group have been determined by single-crystal X-ray diffraction analysis.

TL;DR: Methods that allow for the identification of prenylated and methyl-esterified carboxyl-terminal cysteine residues in proteins radiolabeled in vitro and in vivo are described.

TL;DR: Through radiolabeling methods and use of a cell-free system, the major features of the trypanosome GPI biosynthetic pathway have been elucidated and it is likely that these techniques, successful in the study ofTrypanosomes, will be applicable to other eukaryotic cells.

TL;DR: In this article, the principles of electrostatic levitation where the positioning and stabilization of a sample are accomplished by applying appropriate electrostatic forces to a charged sample are outlined, and attention is focused on a feedback control algorithm, drop-launching method, and four-drop levitator.

TL;DR: These methods are applied to two independent YAC clones encoding the Cu/Zn superoxide dismutase gene (SOD-1) and constructed from these two overlapping clones a restriction enzyme map spanning approximately 600 kb that may prove useful in investigating the genetic basis of Down syndrome.

TL;DR: The mapping strategies, approaches, and protocols for using PFGE in physical mapping complement the classical genetic maps and provide a wider range of markers for mapping.

TL;DR: A collection of methods that have been successfully applied to the construction of long-range maps, including CpG dinucleotides in their recognition sequences and provide landmarks helpful for locating transcribed sequences are presented.

TL;DR: The sequential incorporation of labeled nucleotides into DNA fragments separated on pulsed-field gels allows the identification of the chromosomal origin of replication, the rate and direction of replication fork movement, and the localization of the replication terminus in a single experiment.

TL;DR: Current evidence relevant to this topic is reviewed and experimental methods useful in the study of dynamic fatty acylation are presented, including protocols for metabolic labeling with fatty acids, pulse-chase experiments, linkage analyses, and chemical identification of fatty acids.

TL;DR: The dynamics of single DNA molecules undergoing steady-state and pulsed-field gel electrophoresis have been studied using T2 DNA molecules labeled with acridine orange and visualized with a fluorescence microscope, and it was observed that DNA molecules moving through the gel display an extension-contraction motion.

TL;DR: In this article, over 300 mutants of bacteriophage T4 lysozyme have been constructed and about 80% of these give crystals suitable for X-ray diffraction analysis.

TL;DR: In this article, the authors describe applications of pulsed-field gel electrophoresis (PFGE) techniques to experimental studies with Saccharomyces cerevisiae and give a protocol for preparing chromosomal-sized DNA from yeast.

TL;DR: A simple method for systematically screening crystallization parameters on a pH vs precipitant concentration grid has been used to crystallize several proteins as discussed by the authors, showing that most proteins crystallize over an approximate range of 1.5 to 2 pH units and 20% saturation of ammonium sulfate solutions.

TL;DR: In this paper, the authors describe the production of crystals of monoclonal antibody Fab fragments complexed with influenza virus neuraminidase, and a new technique for photographing protein crystals, using backlighting and an arrangement of colored filters is described.