基因表达差异显示实验

This technique allows specific DNA sequences to be directly localized on chromosomes. The probe DNA is labeled by one of several methods that are described in this chapter. The labeled DNA is then applied onto chromosomes fixed on slides under conditions that allow specific hybridization of t ...

Recent technological advances, not least in somatic cell genetics, have accelerated progress in human genome mapping and reverse genetics. In particular, chromosome mediated gene transfer (CMGT) (1) and irradiation and fusion gene transfer (IFGT) (2) have been exploited to great effe ...

In the early 1980s, a new type of DNA element was described that consisted of arrays of tandemly repeated short sequence units of 9–64 bp (reviewed in ref. 1). A large number of these arrays have subsequently been discovered. These have become known as minisatellites (by analogy with much larger satell ...

DNA fingerprinting without doubt represents one of the most significant advances in forensic science this century. Central to this technology, which is based on the analysis of the genetic component of cells, is the use of DNA probes to regions of the human genome that exhibit great variability b ...

The genetic disorders of hemoglobin, notably, sickle cell anemia and the α- and β-thalassemia, are the commonest genetic diseases in humans. Furthermore, the majority of these mutant globin genes, particularly those causing β-thalassemia, are owing to point mutations that do not involve c ...

Hereditary diseases (1) are diseases that are passed on from one generation to the next. They are caused by one or more genetic defects in consequence of point mutations, small insertions and deletions, or chromosomal rearrangements, notably deletions, duplications, inversions, inser ...

The polymerase chain reaction (PCR) is a rapid method for the amplification and analysis of DNA sequences, and has greatly simplified the identification of mutations leading to genetic diseases (1–3). The exquisite sensitivity of this method can also be exploited to demonstrate the prese ...

The first consistent chromosomal abnormality to be described in a tumor cell was the Philadelphia chromosome (1), but it was not until the advent of high resolution chromosome banding (2,3) that the occurrence of other abnormalities in malignant cells could be fully investigated. Since then, ...

Many standard techniques involving electron microscopy, tissue culture, and protein (antigen) analysis have been developed (reviewed in 1) for virus diagnosis and typing. Since most viral infections can be assigned to a particular virus type by these means, it is now increasingly import ...

Family studies have provided experimental observations enabling geneticists to recognize many human genetic traits and diseases. Single-gene Mendelian traits are usually deduced by straightforward inspection of the data, but sophisticated statistical methods have had to ...

Protocols for the sequence analysis of conventional single-stranded or double-stranded DNA templates are often unsuitable for the direct sequencing of DNA fragments generated by the polymerase chain reaction (PCR) (1,2). The features that can distinguish PCR products as templates ...

The development of techniques for the analysis of specific DNA sequences has led to the discovery of a vast amount of variation of DNA sequence among different individuals. Consequently, it is now usually possible to distinguish the two parental copies of a particular chromosomal region in an i ...

Gene mapping by the analysis of traits segregating in human pedigrees is a major goal of linkage analysis (1), itself firmly rooted in the statistical technique of maximum-likelihood estimation (MLE; 2). The quantity estimated is most often the recombination fraction (ϑ), using the now well- ...

Gene expression in eukaryotes is regulated primarily at the level of transcription. The genomes of higher eukaryotes contain many more genes than are used in any single differentiated cell type, and cell differentiation can be viewed as the result of decisions regarding which genes will be ex ...

Genetic mapping of phenotypic traits in mammals has historically relied on meiotic recombination together with the use of polymorphic markers. Some examples of these markers, in chronological order, are coat color alleles, protein polymorphisms, restriction fragment length po ...

The overall goal in human genetic disease is to understand the biological function of normal genes, and how, once perterbed, they lead to a diseased state. The limitations of using human material for experiments, the ease with which one can work with mice, and the similarities between the two organis ...

Embryo cryopreservation can greatly assist in the management of transgenic mice by providing a low cost means of storing mouse lines or strains while they are not needed (3,12,13,19,27,31). There are several cryopreservation protocols which are very effective for mouse embryos. The cryop ...

Random and selective techniques to alter the mouse genome are now providing powerful tools for biomedical research. These strains provide experimental systems for understanding gene function, for studying defects in specific human genetic diseases, for preclinical testing of t ...

Primate embryonic stem (ES) cells are derived from preimplantation embryos and are capable of prolonged undifferentiated proliferation in culture. Under particular conditions, these cells differentiate into derivatives of endoderm, mesoderm, ectoderm, and trophoblast ( ...

In the ten years since the first gene-targeting experiments were performed in murine embryonic stem (ES) cells (1–5) and the mutations successfully transmitted through the mouse germline (6–11), the application of a variety of gene-targeting methodologies has generated a remarkable n ...