Showing papers in "Methods in 1993"

TL;DR: The two-hybrid system is a yeast-based genetic assay for detecting protein-protein interactions and has been used successfully to analyze interactions of nuclear, cytoplasmic, mitochondrial, and membrane-associated proteins from a variety of organisms.

TL;DR: The numbers of transformants obtained with these protocols are sufficient for complex genomic, cDNA, and gene fusion libraries, facilitating the molecular analysis of the structure and function of genes from higher eukaryotes.

TL;DR: Genetic methods using yeast to analyze a DNA-binding protein to determine the sequence of the DNA sites to which the protein binds and the location of the domain and specific amino acid residues in the protein responsible for DNA binding are described.

TL;DR: Three methods for manipulation of YAC DNA will aid significantly in the construction of physical genome maps and provide powerful genetic tools for the study of genome function.

TL;DR: The features of the Lipofection transfection methodology are described, which is convenient, reproducible, efficient, and effective in a wide variety of tissue culture cell types.

TL;DR: Studies on immunosuppressant mechanisms of action are providing insights into signal transduction in both yeast and T lymphocytes, and the primary drug targets, the immunophilins cyclophilin and FKBP, are highly conserved from yeast to human.

TL;DR: Critical considerations in the design of library screens and routine reporter gene assays are presented, and vectors and protocols that have proven useful for these types of studies are described.

TL;DR: A family of transducing viral vectors that encode antisense RNA targeting critical early events in the replicative cycle of targeted viruses, including human immunodeficiency virus-1 (HIV-1) and herpes simplex virus- 1 (HSV-1), may become powerful analytic and therapeutic tools for the stable introduction of transgenes including antisense genes into cells.

TL;DR: Methods for liposome preparation, screening cell uptake, and sub-cellular fractionation for delivering RNA to cells by liposomes are described.

TL;DR: Protocols for synthesizing high-quality synthetic RNA on an automated DNA/RNA synthesizer are presented, and descriptions of reliable techniques of analysis and purification of synthetic oligoribonucleotides are outlined.

TL;DR: The hairpin ribozyme as discussed by the authors is a catalytic RNA capable of specifically cleaving a variety of substrate RNA sequences in either a cis or a trans reaction, and it has been shown to be a potentially powerful in vivo regulator of gene expression.

TL;DR: General considerations for cloning by complementation in yeast using heterologous cDNA expression libraries are discussed and strategies for complementation of ts and null mutations in essential genes are presented together with transformation and selection protocols.

TL;DR: V vectors for the expression of mammalian receptors in yeast, reporter genes, yeast host strains, and simple assays that monitor receptor transcriptional activity are described.

TL;DR: Methods for using synthetic peptides to specifically probe the molecular mechanisms for calcium-dependent regulation of contraction in cardiac and smooth permeabilized (or skinned) muscle are described and implicate the phosphate release step as the most likely regulatory step in the crossbridge cycle affected by the peptide and, by extension, troponin I.

TL;DR: A method for cloning functional yeast homologs of genes identified in other species is presented here, which allows rapid analysis of protein function using the techniques of yeast genetics.

TL;DR: A mechanism for making cells permeable has been devised to allow an effective examination of both adenylyl cyclase and radioligand binding in an environment that resembles an intact cell.

TL;DR: It is emphasized that small synthetic peptides can mimic the biological activity of their native protein and that small linear sequences are important molecular recognition sites in protein–protein interactions.

TL;DR: Synthetic peptides provided the greatest flexibility and specificity, allowing the precise localization of amino acid sequences of S-antigen required for a particular immunological function such as antibody binding, T-cell proliferative responses, pathogenicity, and the induction of tolerance.

TL;DR: This paper describes the available expression vectors and protein sorting systems, and discusses appropriate strategies for expression of functional mammalian proteins in S. cerevisiae.

TL;DR: These studies provide valuable information to aid in the design of synthetic hormone analogs and for further analysis of the structure–function of hCG via site-directed mutagenesis through systematic approach to the identification of potential receptor binding sites.

TL;DR: This finding suggests that in this context a U5 ribozyme cannot permanently suppress HIV-1 production, and is useful for determining the intracellular efficiency of ribozymes targeted to different sites in the HIV genome.

TL;DR: Screening for interfering activities provides an alternative to complementation as a method of identifying genes from organisms that are not as amenable to genetic analysis as yeast.

TL;DR: Methods for measuring the attachment of viable cells to immobilized peptides to identify sequences possessing cell binding activity are described and two of the 30-mers were found to support cell attachment.

TL;DR: A method for the isolation of cDNAs that encode proteases of known amino acid specificity is presented and is useful for the genetic analysis and identification of inhibitors of proteases whose genes have already been cloned.

TL;DR: The elements necessary for the design, construction, and testing of anti-viral ribozymes in vitro and in vivo are reviewed and some of the elements that must be considered in designing a ribozyme as a potential anti-Viral are described.

TL;DR: In this paper, the role of various regions of Pγ, the amino-terminal, the central cationic, and the carboxylterminal regions, in their interactions with Pαβ and αt* was examined.