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细胞功能测定

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Analysis of DNA Topology in Yeast Chromatin

Topology of closed circular DNA is affected by its packaging into nucleosomes and potentially by alteration of nucleosome structure. Changes in topology that reflect alterations in chromatin structure can be measured and quantified using closed circular plasmids from living yea ...

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Monitoring DNA Breaks in Optically Highlighted Chromatin in Living Cells by Laser Scanning Confocal Microscopy

The recognition and repair of DNA lesions occurs within a chromatin environment. Genetically tagging fluorescent proteins to DNA damage response proteins has provided spatial and temporal details concerning the establishment of biochemical subnuclear regions geared towa ...

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Differential Display Using Random Hexamer-Primed cDNA, Motif Primers, and Agarose Gel Electrophoresis

Polypeptide growth factors stimulate cellular proliferation by binding to the extracellular domain of transmembrane receptors and thereby activating intracellular signal transduction pathways. One cellular response to mitogenic stimulation is the sequential tran ...

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Fingerprinting by Arbitrarily Primed PCR

PCR using primers of arbitrary sequence can generate a reproducible fingerprint of products from DNA (1,2). Differences in the fingerprint of products generated from DNA of related organism are a result of polymorphisms. These polymorphisms proved useful markers for genetic mapping ( ...

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Fluorescent Differential Display

Since their first introduction in 1992 (1,2), differential display (DD) and its relatives have been used extensively in various fields of biology where the identification of differentially expressed messages are of particular interest and importance (for review, see ref. 3). These new te ...

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Cloning Differentially Expressed Genes by Using Differential Display and Subtractive Hybridization

Differential gene expression occurs in all phases of life, including development, maintenance, injury, and death of an organism. Being able to identify these genes will help understand not only gene function but also the underlying molecular mechanisms of a particular biological syst ...

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A Direct-Sequencing-Based Strategy for Identifying and Cloning cDNAs from Differential Display Gels

This report describes an approach to identifying and cloning messages from differential display (DD) gels that has markedly reduced the occurrence of false positives as well as the time required for the process. Most importantly, the use of Northern blots with potentially contaminated p ...

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Direct Sequencing of Differential Display PCR Products

The polymerase chain reaction (PCR)-based mRNA differential display (1) has become an increasingly popular alternative technique for isolating genes of interest in a variety of in vitro and in vivo systems (for review, see ref. 2), as compared to such conventional techniques as differenti ...

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Differential Screening of Differential Display cDNA Products by Reverse Northern

Differential Display (DD) is a widely used methodology for cloning differentially expressed genes. Since its inception in 1992 (1), much effort has been made to improve and optimize the technique (2,3). One major bottle-neck remains, though, in the screening step for the verification of the cDNA ...

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Screening for Positive Clones Generated by Differential Display

Differential display is a powerful method to identify genes that are differentially expressed in different tissues or in the same tissue exposed to different treatments (1,2). The method can be performed with relatively little biological material, and the resulting partial cDNAs can be c ...

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RC4DRestriction Fragment Length Polymorphism-Coupled Domain-Directed Differential Display

Since many life processes, such as development and reproduction, depend on differential gene expression, the spatiotemporal pattern of gene expression is of prime biological interest. Accordingly, the concepts of the mRNA differential display (DD) and RNA fingerprinting techni ...

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Cloning of the 3 Noncoding Regions from Several Members of Heat Shock Protein Gene Families by Differential Display

Several structural and metabolically important proteins in eukaryotes are encoded by multigene families. For understanding the function of each member of a gene family, it is essential to isolate the representative cDNAs encoding the protein of interest. In the past, this has been especi ...

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Identification of ImmediateEarly Gene Targets of the Raf-1 Serine/Threonine Protein Kinase Using an Estradiol-Dependent Fusion Protein, Raf-1:ER

The Raf-1 serine/threonine protein kinase is a central component of the Ras/Raf/MEK/MAP kinase cascade, a highly conserved signaling pathway responsible for the transmission of signals emanating from cell surface growth factor, cytokine, and hormone receptors into the nucleus (1–4 ...

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Isolation of Cytokine-Inducible Genes from Hematopoietic Cells by Differential Display

Hematopoietic stem cells have the capacity to divide and differentiate into several different hematopoietic cell lineages. Hematopoietic growth factors (cytokines) promote both the growth and differentiation of stem cells and committed progenitor cells. Cytokines bind and ...

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Hormone-lnducible Genes in Prostate Cells

Prostate gland development, growth, function and pathology are strongly influenced by steroid hormones. Androgens greatly influence the prostate, but the physiological maintenance of the gland can also depend on the presence of estrogens and other steroid hormones, retinoids and ...

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Application of Differential Display in Studying Developmental Processes

Developmental processes are intricate networks of events by which a fertilized egg proliferates and differentiates into a fully functional multicellular organism (1). Mechanisms are used by dividing cells to create asymmetries in their information content that then allow the eme ...

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Identification of Novel Genes Involved in Adipose Differentiation by Differential Display

In vertebrate animals, adipose tissue functions primarily to store excess energy in the form of triglycerides in periods of nutritional abundance and mobilize it in response to fasting (1,2). It also plays important roles in fatty acid metabolism and glucose homeostasis (3). Recently, adi ...

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Isolation of Song-Regulated Genes in the Brain of Songbirds

We have applied the differential display (DD) technique (1) to isolate genes whose expression is regulated in the brain of songbirds when they hear song of their own species. Song is known to cause a marked increase in mRNA levels of two immediate early genes, ZENK and c-jun, in the auditory forebrain of son ...

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Identification of Vertebrate Circadian Clock-Regulated Genes by Differential Display

Differential display is ideally suited for the identification of genes involved in complex physiological events. Unlike subtractive hybridization techniques, differential display requires no previous knowledge about the dynamics of “important” gene products. Since the o ...

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Corneal Assay for Angiogenesis

In order to develop angiogenic and antiangiogenic strategies, concerted efforts have been made to provide animal models for more quantitative analysis of in vivo angiogenesis. In vivo techniques consist of the cornea pocket and iris implant in the eye, the rabbit ear chamber, the dorsal skin ...

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