遗传学实验技术

The molecular basis of parental imprinting is still unknown, but strong evidence points to DNA methylation as one of the mechanisms involved. Expression of imprinted genes was found to be altered in transgenic mice deficient in DNA methyltransferase 1 (Dnmt1) 1. Recently two new methyltran ...

Introduction of exogenous DNA into Caenorhabditis elegans is important for examining the expression of altered or reporter gene constructs, rescuing mutant genes, and studying gene function in vivo. Until recently, germ-line injection was the most commonly used method for transfo ...

One benefit of the nematode Caenorhabditis elegans as a model system is the ease to conduct forward genetic screens and to isolate mutants with phenotypes of interest. However, identifying the mutated genes requires positional cloning, which can be laborious and time consuming. Inserti ...

WormBase is a comprehensive repository for information on Caenorhabditis elegans and related nematodes. Although the primary web-based interface of WormBase (http://www.wormbase.org/) is familiar to most C. elegans researchers, WormBase also offers powerful data-mining fe ...

Because of technical hurdles, large-scale cell culture methods have not been widely exploited until recently for the study of Caenorhabditis elegans. Culturing differentiated cells from larvae and adult worms is probably not technically feasible because of difficulties in remo ...

Optical methods provide a noninvasive way to monitor calcium transients in Caenorhabditis elegans. Imaging techniques are particularly appealing in C. elegans because worms are optically transparent and can be imaged while fully intact. Furthermore, a variety of genetically enc ...

Whether by patch-clamp techniques or the use of fluorescent vital dyes, measurements of transepithelial ion flux in mammals are limited by cell accessibility. Furthermore, redundant functions and complex regulatory mechanisms can mask loss-of-function phenotypes through co ...

High-pressure freezing (HPF) is capable of converting liquid water, to a depth of approx 0.6 mm, into amorphous ice nearly instantaneously. At midbody, an adult Caenorhabditis elegans hermaphrodite approaches its widest girth of approx 0.1 mm. In theory, an entire living adult animal can be phy ...

The Caenorhabditis elegans gonad is a simple model to investigate molecular mechanisms that regulate fundamental cell and developmental processes. The strength of the model is that C. elegans is amenable to genetic manipulation. The complete genome sequence, advances in mass spectr ...

The nematode Caenorhabditis elegans provides numerous experimental advantages for the identification and characterization of genes required for the function of the nervous system. These advantages include forward and reverse genetic tractability, a relatively simple bo ...

The identification and cloning of the green fluorescent protein (GFP) from jellyfish marks the beginning of a new era of fluorescent reporters. In Caenorhabditis elegans, genetically encoded markers like the fluorescent proteins of the GFP family became the reporter of choice for gene e ...

Caenorhabditis elegans is well suited for toxicological studies owing to its established biology, short generation time, large brood size, and readily scorable life traits. Quantitative parameters of C. elegans that can be assayed include growth, size, progeny production, behavio ...

The genome of the nematode Caenorhabditis elegans was the first animal genome sequenced. Subsequent sequencing of the Caenorhabditis briggsae genome enabled a comparison of the genomes of two nematode species. In this chapter, we describe the methods that we used to compare the C. elegans ge ...

Functional genomic analyses in vertebrate model systems, including fish, frogs, and mice, have greatly contributed to our understanding of embryonic development and human disease. However, new molecular tools and strategies are needed to meet the increasing demands of linking sequ ...

Genetic transformation is an important technology that provides unique opportunities to find, isolate, and analyze genes, as well as to create organisms with unique functional characteristics. Insect biologists have been developing genetic transformation technologies th ...

Advances in genome sciences are demonstrating the dynamic nature of noncoding DNA regions, which are comprised largely of repetitive elements with no apparent function. Retroposons are one class of mobile genetic elements that amplify and move about the genome via a copy-and-paste mech ...

Genome size differences among crop plants are largely due to unequal accumulation of repetitive DNA sequences, mainly transposable elements (TEs). Over the past decade, many families of miniature inverted-repeat transposable elements (MITEs) have been identified and characte ...

In many eukaryotes, the introduction of double-stranded RNA (dsRNA) into cells triggers the degradation of cognate mRNAs through a posttranscriptional gene silencing mechanism. This phenomenon has been called RNA interference or RNAi. Several methods for delivering dsRNA into t ...

Insertions in specific genes belonging to large and homogeneous gene families often do not cause a visible phenotype due to genetic redundancy. Therefore, several single-insertion mutants may have to be combined into double or even triple mutants in order to obtain a loss-of-function phe ...

DNA double-strand breaks provide a powerful means to modify the genome. This chapter describes how to generate and use these breaks to target specific sequences, or other modifications to the Drosophila genome. Both P element dependent gene conversion, in which the chromosomal DNA is broken, ...