遗传学实验技术

For structured RNAs that possess catalytic activity, this activity provides a powerful probe for measuring the progress of folding and the effects of RNA chaperone proteins on the folding rate. The crux of this approach is that only the natively folded RNA is able to perform the catalytic reactio ...

As catalytically active RNAs, ribozymes can be characterized by kinetic measurements similar to classical enzyme kinetics. However, in contrast to standard protein enzymes, for which reactions can usually be started by mixing the enzyme with its substrate, ribozymes are typically s ...

Ribonucleoprotein complexes (RNPs) play crucial roles in a wide range of biological processes. Here, we describe experimental approaches to the UV crosslinking-based identification of protein-binding sites on RNA, using multicomponent Saccharomyces cerevisiae RNPs of the R ...

Large RNA molecules play important roles in all aspects of cellular metabolism ranging from mRNA splicing and protein biosynthesis to regulation of gene expression. In order to correctly perform its function in the cell, an RNA molecule must fold into a complex tertiary structure. Folding of ...

Sequence-specific protein-DNA interactions mediate most regulatory processes underlying gene expression, such as transcriptional regulation by transcription factors (TFs) or chromatin organization. Current knowledge about DNA-binding specificities of TFs is bas ...

RNA-based regulation is increasingly recognized as an important factor shaping the cellular transcriptome. RNA-binding proteins that interact with cis-regulatory motifs within pre-mRNAs determine the fate of their targets. Understanding posttranscriptional networ ...

Protein–protein interactions are vital to all living cells. Many different in vivo and in vitro approaches are published which enable detection and determination of interactions between two proteins. However, most of these techniques are not designed to test for multi-protein inter ...

The analyses of protein-protein interactions are crucial for understanding cellular processes including signal transduction, protein trafficking, and movement. Protein fragment complementation assays are based on the reconstitution of protein function when non-act ...

In plants and all other multicellular organisms, both the intra- and extracellular environments are filled with dynamic biomolecular interactions that control many biological processes. Most of these interactions are biochemical in nature and often exist between proteins. For i ...

Phytohormones are low molecular weight compounds that are produced by plants to regulate growth and development and also in response to biotic and abiotic stresses. The quantitative analysis of these molecules, which is essential for a better understanding of their physiological fun ...

Every biological organism relies for its proper function on interactions between a multitude of molecular entities like RNA, proteins, and metabolites. The comprehensive measurement and the analysis of all these entities would therefore provide the basis for our functional and mec ...

The subcellular energy organelles (chloroplast, mitochondria, and peroxisome) in plants are responsible for major metabolic processes including photosynthesis, photorespiration, oxidative phosphorylation, β-oxidation, and the tricarboxylic acid cycle. Arabid ...

Live cell imaging is an essential methodology for studying the structure, dynamics, and functions of cells in a living plant under normal or stressed growth conditions. Arabidopsis thaliana is perfectly amenable to various live microscopy techniques. In this chapter, we provide guide ...

Flow cytometry, and the accompanying technology of cell sorting, represents an established and valuable experimental platform for the analysis of cellular populations. Applications involving higher plants, which started to emerge around 30 years ago, are now widely employed both ...

Over the past decade, confocal microscopy and the ever-expanding toolchest of fluorescent protein (xFP) markers and technologies have become routine methods for the biological laboratory. A common use of xFP fluorophores is in localizing proteins and the subcellular structures wi ...

The objectives of electron microscopy ultrastructural studies are to examine cellular architecture and relate the cell’s structural machinery to dynamic functional roles. This aspiration is difficult to achieve if specimens have not been adequately preserved in a “living state”; ...

The understanding of cellular and subcellular functions often relies on the ability to visualize proteins as close as possible to their endogenous locations. A number of immunocytochemical techniques have been developed to detect proteins in situ using specific antibodies raised ...

Optical microscopy has developed as an indispensable tool for Arabidopsis cell biology. This is due to the high sensitivity, good spatial resolution, minimal invasiveness, and availability of autofluorescent proteins, which can be specifically fused to a distinct protein of inter ...

Gene activity is regulated via chromatin dynamics in eukaryotes. In plants, alterations of histone modifications are correlated with gene regulation for development, vernalization, and abiotic stress responses. Using ChIP, ChIP-on-chip, and ChIP-seq analyses, the direct bind ...

Chemical genetics utilizes small molecules to perturb biological processes. Unlike conventional genetics methods, which involve the alteration of genetic information mostly with lasting effects, chemical genetics allows temporary and reversible alterations of biolo ...