遗传学实验技术

Characterizing gene function is one of the major challenging tasks in the postgenomic era. To address this challenge, we developed GeneFAS (gene function annotation system), a computer system with a graphical user interface for cellular function prediction by integrating informat ...

Identifying the subcellular compartment of a protein is an important step toward assigning protein function. Starting with a clone containing the open reading frame (ORF) of interest, it is possible to attach a variety of short amino acid tags or fluorescent proteins and detect the location of ...

The era of genomics has opened new possibilities for the computational prediction of protein function. In particular, the comparison of fully sequenced genomes allows us to investigate the so-called genomic context of a gene, which includes its chromosomal positioning relative to oth ...

Small interfering RNAs (siRNAs) have been widely exploited for nucleotide-sequence-specific posttranscriptional gene silencing, as a tool to investigate gene function in eukaryotes, and they hold promise as potential therapeutic agents. Conventionally designed siRNAs a ...

With the completion of the human genome sequencing project in 2001, the identification of novel markers is rapidly gaining importance. It is increasingly recognized that SNPs (single nucleotide polymorphisms) are good markers for disease susceptibility. SNPs are DNA sequence vari ...

Analysis and detection of DNA variation is important in any field of biology. Hence, numerous methods have been developed to analyze DNA. A simple yet effective way of analyzing DNA is by denaturant capillary electrophoresis (DCE). The method is in theory applicable to 95% of the human genome. The me ...

There is a demand for technologies that allow the interrogation of large numbers of SNP polymorphisms, both in whole-genome panels and in smaller custom designed sets, to attempt to elucidate the nature of complex disease through linkage and association studies. The Illumina BeadArray te ...

Recurrent structural alterations, such as amplification, deletion, or translocation, are hallmark features of the cancer genome. Mapping of these DNA copy number aberrations, using approaches such as comparative genomic hybridization (CGH), has enabled the discovery of bona fide ...

As the number of mammalian genomes to be completely sequenced continues to grow, researchers attempting to identify mutations responsible for inherited disease increasingly take advantage of studies undertaken in alternative species, usually humans, to identify and investi ...

With the increasing knowledge on the genetic alterations associated with various cancers, molecular analysis of these alterations on formalin-fixed paraffin-embedded tissue is of increasing importance. Multiplex ligation-dependent probe amplification (MLPA) is a novel ...

Comprehensive analyses of the human epigenome may be of critical importance in understanding the molecular mechanisms of complex diseases, development, aging, tissue specificity, parental origin effects, and sex differences, among other systemic aspects of human biology. How ...

Interactions between cis-acting elements and proteins play a key role in transcriptional regulation of all known organisms. To better understand these interactions, researchers developed a method that couples chromatin immunoprecipitation with microarrays (also known as ...

The yeast artificial chromosomes (YAC) system makes it possible not only to clone large DNA fragments but also to simplify the physical mapping and functional analysis of chromosomes and genomes through YAC manipulation. To manipulate large DNA fragments cloned into YACs, YAC fragment ...

DNA cloned in yeast artificial chromosomes (YACs) is a valuable resource for functional experiments in cell culture as well as whole animal systems. Where the size or chimerism of a YAC clone are limiting factors it may be desirable to generate recombinant YAC clones. One such approach is based on mi ...

Mouse yeast artificial chromosomes (YACs) are useful platforms for manipulation of targeting vector design and construction, particularly in circumstances where polymerase chain reaction-mediated amplification of targeting arms proves fruitless or the cloned DNA is inh ...

The generation of transgenic animals has become an important tool in helping to understand the roles of genes in maintaining health and the roles of human gene variants in contributing to disease susceptibility. In order to model the subtleties of gene expression, which include tissue-spe ...

Genomic-type transgenes are usually expressed in appropriate spatial- and temporal-specific manners. The largest genomic transgenes can be prepared using yeast artificial chromosomes (YACs). Normally, YAC transgenic mice are produced by standard pronuclear microinjec ...

Transgenic mice produced with human yeast artificial chromosomes (YACs) generally display transgene expression patterns that reflect those of the normal human host. Because mice are expensive and time-consuming to generate and maintain, extensive mutation-phenotype corr ...

In addition to the well-known applications of yeast artificial chromosomes (YACs) in classical molecular genetics, they also are used for molecular cytogenetic studies. YACs, as well as other locus-specific probes like DNA, plasmids, cosmids, P1-clones, or bacterial artificial chr ...

This chapter describes the methodology used both in performing the electrophoretic karyotype of the protozoan parasite Trypanosoma cruzi and mapping the genetic markers of the chromosomal bands, the construction of chromosome-specific YAC contigs, and their use to assign a chrom ...