植物学技术

Changes in gene expression mediated by DNA-binding protein factors are a crucial part of many signal transduction pathways. Generally, these regulatory proteins are low abundant and thus their purification and characterisation is labour- and time-intensive. Here we describe a work ...

For understanding the mechanism of transcriptional regulation, it is essential to know which transcription factor is bound in vivo to the promoter to be analysed. If transcription from a given promoter is regulated by developmental or environmental stimuli, the question of inducible v ...

The rapidly increasing amount of entirely sequenced genomes generates a need for fast and efficient methods to elucidate gene functions. Functional complementation of yeast mutants, displaying selectable phenotypes, has been used very successfully in the past years to isolate ma ...

Protein phosphatases act to reverse phosphorylation-related modifications induced by protein kinases. Type 2C protein phosphatases (PP2C) are monomeric Ser/Thr phosphatases that require a metal for their activity and are abundant in prokaryotes and eukaryotes. In plants, such ...

Dynamic and reversible protein–protein interactions have a pivotal function in all living cells. For instance, protein–protein interactions are involved in the assembly and regulation of multimeric enzymes and transcription factors, various signal response pathways, intr ...

Fluorescence cross-correlation spectroscopy (FCCS) is a technique that physically evaluates the molecular interaction between two fluorophore-tagged molecules such as proteins and oligonucleotides in a quantitative manner. Because it simply makes use of the coincident ...

Dynamic networks of protein–protein interactions regulate numerous cellular processes and determine the ability of cells to respond appropriately to environmental stimuli. However, the study of protein complex formation in living plant cells has remained experimentally d ...

In the unicellular flagellated green alga Chlamydomonas reinhardtii several processes are regulated by the circadian clock. To study circadian controlled processes, the cell’s clock is synchronized in a 12 h light–12 h dark cycle (LD12:12) before the cells are released into constant con ...

Eukaryotes control many aspects of growth and development such as cell cycle progression and gene expression through the selective degradation of regulatory proteins by way of the 26S proteasome. Generally, proteasomal degradation requires the poly-ubiquitylation of degrad ...

Each process involved in the generation of plant body form and function is under the control of signals from the exogenous and/or endogenous plant environment. These controls are necessary for adequate plant adjustment to the prevailing conditions, but at the same time they impose the need for ...

Protoplasts of Arabidopsis thaliana have been isolated from a variety of explant sources with varying degree of success (1–3), Most workers have faced problems in achieving a high frequency of sustainable division of protoplasts in liquid culture. The problem can be alleviated to a certain e ...

The mapping of a large number of cloned sequences on yeast artificial chromosome (YAC) clones 1s a srgmficant technical challenge. Although yeast colony lifts are suitable for hybridizations with a limited number of probes, this approach can be difficult to scale up to accommodate hundreds ...

Since the pioneering studies of Klein et al. (1) and Chrtstou et al. (2) bombardment-mediated transformation has become a useful method for dehvermg foretgn genes mto plant cells (3). The process consists of acceleratmg particles to a speed at which they can penetrate the surface of the cell and be inc ...

The transfer of defined DNA segments mto plant cells 1s an established procedure to study regulation of gene expression (1,2). Techmques for direct DNA transfer and transient expression analysis of introduced genes provide a convenient alternative to stable transformation proced ...

Transgenic plants have become a very important tool for the study of gene function. The Agrobacterzum binary transformation system (1) allows the precise transfer of a defined piece of DNA (T‐DNA) to the plant genome (2,3) of many plant species. In this way, any desired DNA sequence can be mtroduced into ...

Genetic transformation and clonal propagation are techniques that play an important role in the identification and characterization of plant genes and their products. The Joint efforts to develop Arabidopszs thaliana as a model for genetic and molecular analysis of higher plants ha ...

Plant genetic transformation was initiated and developed m the 80s thanks to the convergence of constant progress m the protocol of regeneratton from tissue culture, molecular techniques leading to well-expressed marker genes after transfer m plant cells, and the dtverslficatlon of ...

Direct introduction of DNA into plant protoplasts facilitates a rapid analysis of transient gene expression, as well as the generation of stably transformed transgenic plants. Transient gene expression assays performed after DNA transformation permit a comparative analysis of ...

Chromosome walking is a versatile technique applicable to cloning virtually any gene of interest identifiable by mutations1. The prmciple of this approach consists of using a molecular probe (for instance, a restriction fragment length polymorphism marker) mapping near the target ...

Transposon tagging (1) is a method for the isolation of genes, which display a mutant phenotype, Mutants that contain the transposon insert are “tagged” with the transposon and often display the characteristic chimeric or variegated phenotype owmg to transposase-medtated excision. W ...