遗传学实验技术

Different challenge assays have been used to investigate cellular responses following exposure to DNA damaging agents. Our protocol uses X- or γ-rays or ultraviolet light to challenge cells to repair the induced damage, and chromosome aberrations as a biomarker to indicate DNA repair pro ...

A number of ongoing delayed effects have now been described in the progeny of an irradiated cell. These are grouped under the rubric of radiation induced genomic instability. Perhaps the best characterized is the dynamic production of chromosomal rearrangements in some clonally expand ...

Extensive fragmentation of nuclear DNA occurs during apoptosis, and the presence of DNA strand breaks is considered to be a marker of the apoptotic mode of cell death. This chapter describes methods to label in situ DNA strand breaks with fluorochromes for detection by flow or laser scanning cyto ...

The human genome has been almost completely sequenced, and at least 30,000 genes have been found (1). Systematic studies of gene expression patterns by using cDNA microarrays have provided a powerful approach to molecular dissection of cells and tissues by comparing expression levels of te ...

Molecular genetic approaches toward the study of gene function in mice have traditionally relied on either overexpression or gene ablation using transgenic or knockout mouse strategies, respectively. More recently, the knockin approach, which often combines features of both tec ...

The injection of a recombinant adenoviral vector into the tail vein of a mouse results in highly preferential infection of the liver and subsequent liver-specific expression of the genes that are inserted into the adenoviral backbone. These characteristics of systemic adenovirus inj ...

Mouse models play an important role in the elucidation of molecular pathways underlying human disease. Mutations in mouse can be generated by a variety of techniques including those using inducing agents such ionizing radiation or chemicals and those that involve genetic manipulati ...

Ttransgenic and knockout mice often show phenotypes that differ from our expectations. Therefore, it is important to carry out a complete pathologic examination using both macroscopic and microscopic analysis. It should be noted that, apart from (un)expected phenotypes caused dir ...

The mouse has become the most profound model system for investigating the genetics and pathogenetics of human diseases. A huge number of new mouse strains has arisen, and much effort has been expended to increase the number of suitable mouse models. In nearly all animal facilities, the maintenan ...

The application of transgenesis has increased exponentially since its introduction in the early 1980s, and it is still one of the most powerful methods to study gene function. As approaches to solve scientific problems became more complex, transgene design evolved alongside. At this mome ...

With the tetracycline (Tc)-controlled gene expression system (Tet system), an experimental approach has been developed that permits alteration of individual gene activities in complex genetic systems in a truly conditional fashion, i.e., it is temporally defined, quantitative, a ...

The use of large genomic clones for transgenesis is necessary when the gene to be expressed is too large to be accommodated in a plasmid-based vector. Large insert clones can be produced with the P1 bacteriophage (P1) (1), P1 artificial chromosome (PAC) (2), bacterial artificial chromosome (BAC) (3 ...

Gene targeting in mouse embryonic stem cells is now a well-established technique that is widely used to create animal models for human disease or to study gene function at the level of the whole animal. The purpose of this chapter is to provide a detailed protocol to investigators without previous ex ...

Reporter molecules are commonly used in transgenic mice to follow in vivo gene expression patterns through all developmental stages of the life cycle in all tissues. Numerous reporters are available for use in transgenic mice. The selection of a particular marker for gene expression will af ...

AMPS (Alignment of Multiple Protein Sequences) is a set of programs for the multiple alignment of protein sequences. The programs build on the sequence comparison technique introduced in 1970 by Needleman and Wunsch (1). The Needleman and Wunsch algorithm when supplied with two protein seq ...

Alloimmunization to the blood group antibody anti-RhD (anti-D) is the most common cause of hemolytic disease of the fetus and newborn. Knowledge of fetal D type in women with anti-D makes management of the pregnancy much easier and avoids unnecessary procedures in those women with a D-negative fe ...

Elucidation of the mechanisms of recognition of mispaired DNA by MutS and subsequent repair by the multicomponent mismatch repair machinery presents a fascinating challenge (1). ATP binding or hydrolysis are implicated in several steps of the process, including enabling MutS to diss ...

Cellular and organism aging have been correlated with accumulated DNA damage (1,2). 8-oxo-7,8-dihydrodeoxyguanine (8-oxoG or GO) is one of the most stable products of oxidative DNA damage. The formation of GO in DNA, if not repaired, can lead to misincorporation of A opposite to the GO lesion and resu ...

Oxidative damage produced by endogenously and exogenously generated reactive oxygen species (ROS) has been implicated in mutagenesis and carcinogenesis and may play an important role in the pathogenesis of aging (1). Among ROS, the hydroxyl radical is highly reactive, producing a var ...

One of the main determining factors for maintaining the informational integrity of the DNA genomes in all organisms is the efficiency of repair of DNA lesions. DNArepair mechanisms have evolved to counteract the deleterious effects of DNA damage. One such repair mechanism is Base Excision R ...