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基因表达差异显示实验

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Molecular Recognition of Diverse Ligands by T-Cell Receptors

T-cell receptors (TCRs) are structurally related to antibodies, and also interact with a diverse set of ligands. TCRs recognize foreign peptide antigens displayed by major histocompatibility complex (MHC) molecules and foreign lipid-based antigens presented by CD1. These inter ...

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Identification of Human MHC Class I Binding Peptides using the iTOPIA Epitope Discovery System

CD8+ T cells recognize antigenic peptides presented in the context of MHC class I. They play a key role in cellular immunity and are crucial for longterm protective immunity to many infectious diseases. The quest for new and enhanced vaccines requires improved means for identification of rele ...

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T-Cell Epitope Mapping in Mycobacterium tuberculosis Using PepMixes Created by Micro-Scale SPOT Synthesis

Mycobacterium tuberculosis (Mtb) remains a major threat to human health worldwide. Although treatment of infection is an important part of tuberculosis control, an improved vaccine is essential for the elimination of this disease. Control of infection with Mtb is dependent on the cellu ...

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High-Throughput T-Cell Epitope Discovery Through MHC Peptide Exchange

Recombinant major histocompatibility complex (MHC) class I molecules complexed with pathogen-specific or other disease-associated antigens have become essential reagents for the analysis of adaptive T-cell responses. However, conventional techniques for the product ...

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T-Cell Epitope Processing (The Epitope Flanking Regions Matter)

Epitopes presented by major histocompatibility complex (MHC) class I molecules for cytotoxic T-lymphocyte (CTL) recognition are derived mainly from cytosolic proteins. Antigen presentation on the cell surface requires correct processing of epitopes by the proteasome, cyto ...

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Identification of MHC Class II Binding Peptides: Microarray and Soluble MHC Class II Molecules

CD4+ T-helper cells recognize antigenic peptides presented by MHC class II molecules. The binding of the nominal peptide to the MHC class II allele is dependent on the amino acid sequence of the peptide as well as on amino acid (aa) residues in the peptide binding groove of the MHC class II allele. MHC class II a ...

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T-Cell Epitope Mapping

Identification of epitopes defined by T-cell responses aids to (1) monitor antigen-specific cellular immune responses (2) guide rational vaccine design, and (3) understand the nature of protective or harmful T-cell responses in diseases with defined target antigens. The 6-h intrace ...

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Identification and Validation of T-Cell Epitopes Using the IFN- EliSpot Assay

The quest for new and enhanced vaccines requires improved means for identification of relevant antigens and the epitopes present within these. While CD8+ T cells recognize antigenic peptides presented in the context of MHC Class I, and play a key role in cellular immunity, CD4+ helper T-cells rec ...

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Pyrosequencing Genotype Storage Techniques

Data storage and data coordination are important aspects of project design and execution. Pyrosequencing� technology allows thousands of data-points to be collected per day. Consequently, a consistent and reliable method of data input and storage is vital. This chapter discusses the s ...

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The History of Pyrosequencing

One late afternoon in the beginning of January 1986, bicycling from the lab over the hill to the small village of Fullbourn, the idea for an alternative DNA sequencing technique came to my mind. The basic concept was to follow the activity of DNA polymerase during nucleotide incorporation into a DNA st ...

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Pyrosequencing Applications

Genetics research has benefited tremendously from the release of the human genome sequence. Subsequent technology has been developed and adapted to accommodate the need for faster, easier throughput of genetic assays. Pyrosequencing� is a unique system that allows the analysis of gen ...

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Web-Based Primer Design Software for Genome-Scale Genotyping by Pyrosequencing

Design of locus-specific primers for use during genetic analysis requires combining information from multiple sources and can be a time-consuming process when validating large numbers of assays. Data warehousing of genomic DNA sequences and genetic variations when coupled with s ...

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Pyrosequencing: A Simple Method for Accurate Genotyping

Pharmacogenetic research benefits first-hand from the abundance of information provided by the completion of the Human Genome Project. With such a tremendous amount of data available comes an explosion of genotyping methods. Pyrosequencing� is one of the most thorough yet simple meth ...

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Universal Primer Applications for Pyrosequencing

Pyrosequencing� is a high-throughput technique for single-nucleotide polymorphism (SNP) genotyping, DNA sequencing, and SNP allele frequency and DNA methylation assays. The Pyrosequencing assay involves initially purifying PCR-generated single-strand DNA labeled w ...

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Multiplex Pyrosequencing for DNA Variation Analysis

Pyrosequencing� technique has been widely used to perform both single-nucleotide polymorphism detection and quantitative DNA methylation analysis. Simplex Pyrosequencing is sufficient to interrogate more than one polymorphic site if these gene variants are within the rea ...

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Allele Quantification and DNA Pooling Methods

Studies utilizing differences in single-nucleotide polymorphism allele frequencies between cases and controls have been widely used in genetic analyses to locate putative genes or chromosomal regions that may be associated with a disease. In these studies the assessment of allele ...

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Analysis of Gene-Specific DNA Methylation Patterns by Pyrosequencing Technology

As the sequence of the human genome is now nearly finished, genome research turns to elucidate gene function and regulation. DNA methylation is of particular importance for gene regulation and is strongly implicated in the pathogenesis of various diseases. The real-time luminometric de ...

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Detecting Mutations That Confer Oxazolidinone Resistance in Gram-Positive Bacteria

Resistance to oxazolidinone antibiotics, including linezolid, in Gram-positive bacteria is mediated by single-nucleotide polymorphisms (SNPs) in the 23S ribosomal RNA. A G2576U change (encoded by a G2576T mutation in the rRNA genes) is found in most resistant clinical isolates of en ...

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Energy Systems for ATP Regeneration in Cell-Free Protein Synthesis Reactions

Supplying energy for cell-free protein synthesis reactions is one of the biggest challenges to the success of these systems. Oftentimes, short reaction duration is attributed to an unstable energy source. Traditional cell-free reactions use a compound with a high-energy phosphate bo ...

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SIMPLEX: Single-Molecule PCR-Linked In Vitro Expression: A Novel Method for High-Throughput Construction and Screening of Protein Libraries

A novel strategy for construction of protein libraries called “SIMPLEX: single-molecule PCR-linked in vitro expression” is described. A pool of genes is prepared and thereafter extensively diluted to give one molecule of DNA per well. Each individual molecule is separately amplified by ...

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