细胞功能测定

Whole-genome radiation hybrid (RH) mapping has proven to be a powerful tool for mapping genes and comparing genome architecture. We describe a protocol for constructing RH panels by rescuing irradiated fibroblast donor cells of any mammalian species by polyethylene glycol fusion to a th ...

Radiation hybrid (RH) mapping has become one of the most well-established techniques for economically and efficiently navigating genomes of interest. The success of the technique relies on random chromosome breakage of a target genome, which is then captured by recipient cells missing a ...

Whole-genome bacterial artificial chromosome (BAC) libraries for ∼40 representative species of mammals provide unique resources for the construction of high-resolution ordered genomic maps, the development of species genetics, and multispecies genome comparisons. Her ...

Phylogenomics research integrating established principles of systematic biology and taking advantage of the wealth of DNA sequences being generated by genome science holds promise for answering long-standing evolutionary questions with orders of magnitude more primary d ...

Comparative genomics is a powerful approach for inferring the history and function of genomic sequence. The generation of bacterial artificial chromosome (BAC)-based physical maps is a proven method for the targeted comparative genomic analysis of genes or regions of interest. ‘Uni ...

The UC Santa Cruz Genome Browser provides a number of resources that can be used for phylogenomic studies, including (1) whole-genome sequence data from a number of vertebrate species, (2) pairwise alignments of the human genome sequence to a number of other vertebrate genome, (3) a simultaneous ...

Multicomponent RNA-protein complexes are essential for eukaryotic gene expression. Some, like the spliceosome, have been studied successfully in vitro using biochemical and structural approaches, but many have not been reconstituted in cell-free systems. Nucleotide analog ...

Ultraviolet (UV)-dependent photochemical crosslinking is a powerful approach that can be used for the identification of RNA-protein interactions. Although 8-azidoATP (8-N3ATP) has been widely used to elucidate the ATP binding site of a variety of proteins, its inability to serve as an ef ...

A large number of proteins contain multiple RNA recognition motifs (RRMs). How multiple RRMs contribute to RNA recognition in solution is, however, poorly understood. Here, we describe a simple biochemical approach called CLAMP (crosslinking and mapping of protein domain) to identify ...

Characterization of novel RNA-protein interactions often demands physical mapping of the RNA binding sites in the protein. This can sometimes be accomplished using radioactively labeled RNA in covalent RNA-protein crosslinking experiments. The position of the radioactive la ...

Mass spectrometry is a powerful tool for the analysis of biomolecules, proteins, nucleic acids, carbohydrates, lipids. In combination with genome sequences that are available in the databases, it has proven to be the most straightforward and sensitive technique for the sequence analys ...

We present a high-resolution mass spectrometric footprinting approach enabling the identification of amino acids in the protein of interest interacting with cognate RNA. This approach is particularly attractive for studying large nucleoprotein complexes that are less amena ...

Oxidative stress is recognized to be involved in many pathological conditions, such as inflammation, arteriosclerosis, and neurodegenerative diseases. Reactive oxygen species (ROS), which induce oxidative stress responses, appear to induce different biochemical and cel ...

Human ribonucleotide reductase (RR) small subunits, M2 and P53R2, play key roles in forming RR holoenzyme and supplying nucleotide precursors for DNA replication and repair. Currently, we are studying the redox property, structure, and function of hRRM2 and p53R2. In the cell-free system, p ...

Antioxidants are health beneficial compounds through their combat with reactive oxygen and nitrogen species and free radicals that may cause tissue damage leading to various diseases. This work reports the development of a simple and widely applicable antioxidant capacity index f ...

Methodology for deriving quantitative structure-activity relationship (QSAR) models based on computed molecular descriptors, representing numerically structural features of polyphenols, and applicable to the antioxidant activity of polyphenols is delineated. T ...

Immunohistochemistry is an important tool that is often used for the diagnosis of several diseases in the pathology laboratory. The quality and sensitivity of immunohistochemical staining is affected by formalin fixation, which results in variable loss of antigenicity, known as a ma ...

As organisms designed to depend upon oxygen to sustain life, humans are necessarily and continually exposed to damaging oxidizing agents. As a vital protective measure, oxygen-dependent organisms have developed a highly evolutionarily conserved mechanism for preventing oxid ...

This chapter describes the use of reversed-phase HPLC with multichannel coulometric electrochemical detection for the routine, sensitive, and simultaneous measurement of oxidized and reduced CoQ10 and CoQ9 in human plasma and serum. Analytes are first resolved chromatograph ...

Antioxidant liposomes provide a unique means of delivering both water and/or lipid soluble antioxidants to tissues thereby affecting disease states or signal transduction pathways modulated by oxidative stress. Considerable evidence suggests that liposome-encapsula ...