遗传学实验技术

Exon trapping (Fig. 1) is a technique that has been developed to identify genes in cloned eukaryotic DNA (1–7). Compared with other techniques for gene identification, exon trapping has two main characteristic features. First, it is independent of the availability of an RNA sample in which the gene ...

Over the last 7 years, there have been enormous changes to the methods being used to sequence DNA. These changes include the increased use of fluorescent dyes to label DNA in the sequencing reaction, improvements to automated sequencers, and the use of high-throughput methods for preparing and s ...

Despite the fact that the genetic blueprint in every cell is identical, an organism is composed of a multitude of different cell types. This cell type complexity or phenotypic differences are the result of differential expression of identical genes. For example, a normal cell expresses a diffe ...

The major goal of the Human Genome Project must be to identify, sequence, characterize and assign specific function to all the genes spread through our 3000 Mb of haploid DNA. Owing to the uniformity and simplicity of the DNA code, it is not an easy task to identify genes even after the region in which they lie has b ...

You have, we hope, come to this chapter wishing to know how to use bioinformatics to locate genes in genomic sequences. You probably want to know which are the easiest and best tools to use, to treat them largely as black boxes, and to know how to assess the likely accuracy of the results you obtain. If so, you have come to t ...

The mapping of genes underlying either simple mendelian or complex traits can be broken down into a number of distinct stages—initial detection of the locus in a genome scan, determination of the most likely map location for the gene, and finally fine mapping of the locus. A number of experimental str ...

Representational difference analysis of cDNA (cDNA RDA) is designed to compare two different mRNA populations resulting in the isolation of differentially expressed genes. RDA was originally developed for genomic DNA as a method to isolate the differences between two complex genom ...

The genome of a given organism is considered in biology as the fundamental invariant (1). It is virtually the same throughout lifetime and, to a lesser extent, over generations. By contrast, genetic information is expressed in complex and ever-changing temporal and spatial patterns throug ...

Profiling the expression status at the level of transcription is a starting point for delineation of the function of both known and unknown gene products. In addition, the identification of genes expressed in a regionally, temporally, or environmentally specific manner is fundamental to ...

As the Human Genome Project nears the completion of the first human sequence, the next great challenge is to elucidate the function of these genes. One route of exploring the function of a gene is by determining its pattern of expression. Various methods are available for detecting and quantitati ...

Functional analysis of the mammalian genome will be a major task of biologic science in the future. Gene trap is a method designed to provide functional information for novel genes (1–3). It offers tagging of a gene (facilitating subsequent cloning), analysis of the expression pattern by a simple s ...

Proteins mediate virtually all biological processes. Understanding the mechanisms by which proteins function requires a knowledge of their three-dimensional (3D) structures. As a consequence of the genome and full-length cDNA sequencing projects, there are several orders of ma ...

A decade ago the first alteration of a gene using embryonic stem (ES) cells was performed in the mouse (1). Since then ES cells have become a powerful tool to generate mutant mice for the analysis of gene function. These mutant mice (also called knockout mice) have provided new insights into normal and path ...

As the completion of genome sequencing efforts leads to the definition of increasing numbers of genes, the need to reliably assign function to identified coding sequences becomes paramount. One means of gaining initial insight into the function of an undefined protein is to develop a map of oth ...

Gene therapy vectors based on adeno-associated virus (AAV) are being used to successfully transduce a number of different tissues, including muscle (1). The first demonstration of muscle transduction by recombinant AAV (rAAV) was reported by Xiao et al. (2) in 1996. In that report, LacZ express ...

An obligatory part of the life cycle of retroviruses (Fig. 1) is a stable, chromosomally integrated form of the virus, known as the provirus. The existence of the proviral form of retroviruses has provided one of the main driving forces for their use as vectors for gene transfer because, in the instance of a r ...

One of the challenges of medical research today is to find ways of bringing our genetic knowledge of cancer to clinical application and to develop improved therapies by exploiting gene-based strategies. By offering increased specificity and reduced toxicity, gene-based approaches p ...

The analysis of traits and disorders that exhibit a straightforward Mendelian genetics, based on the kind of major gene models that are easy to set up in computer programs such as LINKAGE (1), has been enormously successful in facilitating identification of the genes responsible. These monog ...

One of the major tasks in human genome analysis is the identification and typing of DNA sequence variations (1). There are many types of sequence variations in the human genome. One type comprises sequences with variations in the number of repeat units such as short tandem repeat polymorphisms in t ...

The completely unexpected discovery that the RNA molecule has catalytic properties (1,2) has led to a plethora of interest in the identification and utilization of a variety of catalytic RNA molecules, or ribozymes, that occur in nature. Among others, hammerhead ribozyme is a small catalytic ...