遗传学实验技术

Aggregation of repeat-containing proteins is associated with neurodegenerative disorders; a specific example is the established link between expansion of the polyglutamine domain in huntingtin and the appearance of nuclear inclusions in Huntington’s disease. This connec ...

The accumulation of specific aggregation-prone proteins during aging is thought to be involved in several diseases, most notably Alzheimer’s and Parkinson’s disease as well as polyglutamine expansion disorders such as Huntington’s disease. Caenorhabditis elegans disease mo ...

Expansion of repeat sequences beyond a pathogenic threshold is the cause of a series of dominantly inherited neurodegenerative diseases that includes Huntington’s disease, several spinocerebellar ataxias, and myotonic dystrophy types 1 and 2. Expansion of repeat sequences occ ...

A pathological hallmark of many protein-misfolding diseases is the formation of insoluble aggregates. Quantitative methods are needed to better resolve and define the formation, aggregation, and temporal dynamics of soluble misfolded proteins in native settings. In this book ch ...

Protein misfolding is associated with many neurodegenerative diseases, including neurodegenerative diseases caused by polyglutamine expansion proteins, such as Huntington’s disease. The model organism baker’s yeast (Saccharomyces cerevisiae) has provided import ...

Disease-causing polyalanine (PA) expansion mutations have been identified in nine genes, eight of which encode transcription factors (TFs) with important roles in development. In vitro and cell overexpression studies have shown that expanded PA tracts result in protein misfoldi ...

Generation of fusion proteins is a routine procedure in an increasing number of laboratories worldwide. Generally, the cDNA sequence of the protein under study is subcloned in-frame into a vector containing the sequence of a wellestablished epitope. This procedure, although simple and ...

Site-directed mutagenesis (SDM) is a powerful tool for analyze protein structure and function, protein folding, and enzyme mechanism (1). Several protocols for SDM by polymerase chain reaction (PCR) are published (2–8). Here, two protocols, which turned out to be robust and efficient in the au ...

Generating mutant proteins has become a routine strategy for molecular and structural biologists, to understand the structure-function relationship of a given protein of interest. In this, site-directed mutagenesis has proven to be a valuable technique. After the advent of polymer ...

Acinetobacter strain ADP1 (also known as strain BD413) is unusual among bacteria in the frequency with which it incorporates DNA into its chromosome (chr) by natural transformation. Since there is no requirement for uptake sequences to be present in the donor DNA, strain ADP1 can be transformed ...

The study of bacterial pathogenicity in vitro has identified many signals, at the molecular and cellular levels that affect expression of virulence and other factors in causing disease. Because these pathways are not necessarily reproduced in vitro, their implication and their regul ...

For many biologists, the polymerase chain reaction (PCR) has become an indispensable tool serving many diverse applications, from simple DNA amplification to complex diagnostics. In basic research, a very important application has been the use of PCR in the introduction of site-specif ...

Scanning mutagenesis strategies have proven to be a useful approach to structure-function and protein evolution studies (1), yet great effort is required to generate comprehensive data for a given protein. In these experiments, one or several amino acid residues are changed (i.e., to alani ...

Site-directed in vitro mutagenesis is a powerful tool for investigating the structure-function relationships of proteins and the expression of genes. Often, it is desirable to subject the protein or gene of interest to scanning mutagenesis. The genetic analysis of scores of single or mul ...

Mutagenesis of large plasmids, such as bacterial artificial chromosomes (BACs) or P1-based artificial chromosomes (PACs), can be difficult for various reasons. Because of their large size (up to 300 kbp), unique restriction enzyme sites are usually not available. Even if they are, modific ...

Random mutagenesis of DNA has been an essential tool for investigation of bacteria, fungi, and other organisms. Given the recent explosion in genomic sequence information (e.g., over the past 5 yr there have been 30 published microbial genomes and over 100 microbial genome sequencing projec ...

Site-specific mutagenesis is a powerful tool in molecular biology research. A number of techniques are available today for carrying out site-directed mutagenesis (SDM). Common among them is the oligonucleotide-directed mutagenesis (1). Three widely used procedures, which are ba ...

The introduction of subtle mutations into genomic or cDNA regions of interest provides a powerful approach to the study of gene expression and function. Such manipulations of DNA permit numerous small physical changes, and their effects, to be examined. Unfortunately, most mutagenesis ...

Technological advancement in molecular biology set in motion the directed evolution of the properties of biomolecules, such as protein and nucleic acids. Toward the improvement of the properties in general, events in nature are imitated. The approach, commonly called “directed evol ...

Arabidopsis thaliana is now recognized as a model organism for research in plant biology, especially genetics and molecular biology, and is used for the functional analyses of various genes. However, genomic DNA sequence data reveal that a large number of Arabidopsis genes remain functio ...