遗传学实验技术

Microarray technology can rapidly generate large databases of gene expression profiles. Our laboratory has applied these techniques to analyze differential gene expression in cardiac tissue and cells based on mouse heart transplantation. We have analyzed the different gene exp ...

Large-scale microarray expression profiling studies have helped us to understand basic biological processes and to classify and predict the prognosis of cancers; they have also accelerated the identification of new drug targets. Affymetrix GeneChip� probe arrays are high-dens ...

Serial analysis of gene expression (SAGE), a functional genomics technique, can be used for global profiling of gene transcripts. It relies on the preparation and sequencing of cDNA concatemers, but it does not require prior knowledge of the genes to be assayed (as with microarrays). Once analy ...

The regulation of myocardial gene expression is highly sensitive to any extracellular or intracellular stimulus that affects contractile function. Subtractive suppression hybridization represents a large-scale, unbiased method for detecting transcriptionally and ...

By providing genome-scale information on gene expression, microarray technology has gained popularity in diverse areas including clinical medicine. However, the analysis and interpretation of microarray data are often complicated. This chapter describes various strate ...

Two methods applicable to the measurement of cardiac gene expression by reverse transcription polymerase chain reaction (RT-PCR) are presented. Each method describes a specific technique and includes information on how to optimize the experiments and how to trouble-shoot any prob ...

Quantitative (real-time) PCR (qPCR) represents a highly sensitive, sequence-specific, and reproducible technique for the gel-free detection and quantitation of nucleic acids. Owing to its large dynamic range and throughput, this approach has become the chosen method for rapid qua ...

Quantifying the level of mRNA is central to the study of mammalian gene expression. Conventional approaches such as Northern blotting are often prone to low sensitivity and reproducibility. The RNase protection assay (RPA) provides a sensitive alternative for the detection and quant ...

To understand how cardiac gene expression is regulated, the identification and characterization of cis-regulatory elements and their trans-acting factors by gel mobility shift assay (GMSA) or gel retardation assay are essential and common steps. In addition to providing a general p ...

In situ hybridization allows the detection of specific gene transcripts in tissues, cells or, chromosomes. In the cardiovascular field, this powerful and rapid methodology provides precious insights into the complex gene organization and expression within an heterogeneous cell ...

Forcing ubiquitous expression of a given gene in transgenic Drosophila has become a powerful tool of molecular genetics. Gene misexpression usually causes a dominant phenotype and one may deduce potential functions for the gene from an analysis of this phenotype. Evidently, when a domin ...

The ability to manipulate genes in mammals is providing insights into most aspects of modern biology, including the regulation and function of genes, the mechanisms of developmental and pathological processes, and the generation of animal models for human disorders. Furthermore, the ...

Several methods are available to accomplish gene transfer into mammalian cells, plant cells, yeast, and other organisms. Three of the most promising DNA delivery systems are electroporation (1,2), DNA-coated microprojectiles (3), and microinjection (4,5). Electroporation invo ...

As a result of morphological and developmental characteristics of the early avian embryo, several approaches, taking place at early stages, have been investigated to introduce foreign DNA into the avian genome. They include:

Insertion of foreign DNA into the germline requires access to the chromosomes of cells that give rise to sperm or eggs. From a theoretical point of view, the newly fertilized zygote is the most appropriate cell for genetic alteration because any modification of the genome will be inherited by every c ...

Most common strategies for gene transfer in plants derive from the fact that Agrobacterium tumefaciens contains Ti plasmids which facilitate transfer of T-DNA from the plasmid genome into plant cells with attendant integration of the T-DNA region containing foreign genes into the plant ...

A crucial step in the characterization of recombinant genes is their introduction into a suitable host. Numerous techniques have been developed for introducing recombinant genes into plant cells (see other chapters in this volume, or vol. 6 in this series) (1), one of the most versatile and gener ...

The evolved gene transfer mechanism of Agrobacterium tumefaciens is the transformation method of choice. Its use is simple, gene transfers are precise and result in permanent genetic changes. The range of species known to be infected by this organism has grown from an original group of dicot pl ...

Microalgae are single-celled or colonial photosynthetic organisms that are receiving increasing attention for use their potential usefulness in a number of industrial applications, including bioremediation and the production of high-value specialty chemicals and fuels. ...

Determination of a gene’s transcriptional start site underlies the identification of the proximal promoter region and thus facilitates the subsequent analysis of components controlling its expression, namely, cis-acting regulatory elements and their cognate binding prot ...