遗传学实验技术

Microarray-based screening technologies have revealed a larger than expected diversity of gene expression profiles for many cells, tissues, and organisms. The complexity of RNA species, defined by their molecular structure, represents a major new development in biology. RNA not on ...

Many elegant methodologies have been devised to explore RNA-protein as well as RNA–RNA interactions. Although the characterization of messages targeted by a specific RNA-binding protein (RBP) has been accelerated by the application of microarray technologies, reliable methods ...

The study of recombination in Saccharomyces cerevisiae benefits from the availability of assay systems that select for recombinants, allowing the study of spontaneous events that represent natural assaults on the genome. However, the rarity of such spontaneous recombination re ...

Recombination is involved in many important biological processes including DNA repair, gene expression, and generation of genetic diversity. Recombination must be carefully regulated so as to prevent the deleterious consequences that may result from rearrangements between ...

Because of the availability of the complete sequence of the genome of the model plant Arabidopsis and of insertion mutants for most genes in public mutant collections, the elucidation of the particular role of different factors involved in DNA recombination and repair processes, an import ...

Eukaryotes repair DNA double-strand breaks (DSBs) by homologous recombination (HR) or by nonhomologous end-joining (NHEJ). DSBs are a natural consequence of DNA metabolism, occurring, for example, during DNA replication and meiosis. DSBs are also induced by chemicals and radiation. I ...

African trypanosomes, such as Trypanosoma brucei, are protozoan parasites of mammals that were first described over 100 hundred years ago. They have long been the subjects of biological investigation, which has yielded insights into a number of fundamental, as well as novel, cellular proc ...

The goal of understanding the function of all mammalian genes is best accomplished through mutational analyses. Although the sequence of the mouse genome is now available and many genes have been identified, it is not possible to ascribe functions accurately to these genes in silico. Gene tar ...

The yeast DEL assay is a simple, rapid method for measuring the frequency of reversion of a disrupted his3 gene by homologous intrachromosomal recombination. Reversion to histidine prototrophy results in deletion (DEL) of the disrupting sequence. The DEL assay has been used to study the effe ...

Large-scale genomic rearrangements such as DNA deletions play a role in the etiology of cancer. The frequency of DNA deletions can be elevated by exposure to carcinogens or by mutations in genes involved in the maintenance of genomic integrity. The in vivo DNA deletion assay allows a visual detec ...

The ability to make specific genomic alterations is an invaluable tool to researchers who use genetics and biochemistry to study problems in biology. We have investigated some of the parameters governing DNA fragment transplacement in two commonly used strains of Saccharomyces cere ...

In recent years, triplex-forming oligonucleotides (TFOs) have emerged as powerful tools for site-specific gene modification. Their sequence specificity, binding affinity, and ability to provoke repair and recombination make them promising reagents for altering gene expre ...

In essentially all organisms, double-strand breaks in chromosomal DNA stimulate repair by multiple mechanisms, including homologous recombination. It is possible to use site-specific reagents to produce a break or other recombinagenic damage at a unique site, which makes possible ...

Plasmid DNA transfected into mammalian cells can integrate into mammalian chromosomes by homologous recombination. This phenomenon, known as gene targeting, can be used as a tool to investigate mammalian homologous recombination mechanisms. The chromosomal immunoglobulin ...

Targeted nucleotide exchange (TNE) is a process in which an oligonucleotide bearing sequence complementarity aligns with the sequence of a target gene and directs the alteration of a single base. This technique can be used to repair a point mutation or mediate site-specific mutagenesis. A cr ...

Chromatin immunoprecipitation is a technique that allows one to examine the in vivo localization of proteins to DNA. This technique is well suited for studying genetic recombination since it can provide both a temporal and spatial assessment of the dynamic association of proteins with DNA ...

Holliday junctions are central intermediates in the process of genetic recombination; they form as a consequence of a reciprocal exchange of strands between paired DNA molecules. Enzymes that specifically recognize and process these junctions are necessary for the formation of rec ...

In the search for methods to explore the interaction between proteins and DNA, a plethora of footprinting techniques have been developed, many of which are discussed elsewhere in the present work. Most footprinting techniques are based on the simple premise of specific DNA regions being pro ...

Many studies have been conducted with the aim to stimulate a therapeutic immune response against tumors. In most cases, efforts have been directed toward the induction of tumor-specific cytotoxic T lymphocytes (CTL), because this T lymphocyte subpopulation is considered to play a major r ...

Vaccination of fish against many different pathogenic organisms has made it possible to rear Atlantic salmon in net pen cages and produce fish commercially around the world. In fact, vaccine use is critical for the continued growth of the aquaculture industry and researchers are continual ...