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丁香实验推荐阅读
Use of Next Generation Sequencing (NGS) Technologies for the Genome-Wide Detection of Transposition

Plant transposable elements are ubiquitous in eukaryotes. Their propensity to densely populate the genomes of many plants and animal species has put them in the focus of both structural and functional genomics. Although a number of bioinformatic software have been recently developed f ...

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Massive Indexed Parallel Identification of Transposon Flanking Sequences

The large scale sequencing of insertion element flanking sequences has revolutionized reverse genetics in plant research: Insertion mutants can now simply be identified in silico by BLAST searching the resulting flanking sequence databases. The development of next-generati ...

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Transposon Display: A Versatile Method for Transposon Tagging

Transposon tagging has been used successfully in a range of organisms for the cloning of mutants of interest. In species containing high copy numbers of transposable elements combined with a high transposition rate, forward cloning can be quite challenging and requires specific high-r ...

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Identification and Applications of the Petunia Class II Act1/dTph1 Transposable Element System

Transposable genetic elements are considered to be ubiquitous. Despite this, their mutagenic capacity has been exploited in only a few species. The main plant species are maize, Antirrhinum, and Petunia. Representatives of all three major groups of class II elements, viz., hAT-, CACTA- and Mut ...

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Reverse Genetics in Rice Using Tos17

Transposon of Oryza sativa 17 (Tos17), a Ty1-Copia Class I retroelement, is one of the few active retroelements identified in rice, the main cereal crop of human consumption and the model genome for cereals. Tos17 exists in two copies in the standard Nipponbare japonica genome (n = 12 and 379 Mb). Tos17 cop ...

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Activation Tagging Using the Maize En-I Transposon System for the Identification of Abiotic Stress Resistance Genes in Arabidopsis

Activation tagging is a high-throughput method of overexpressing genes by using an enhancer present in insertion sequences that are randomly inserted in the genome to enhance the expression of adjacent genes. Gain-of-function approaches are advantageous to identify the functions ...

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Molecular Genetics and Epigenetics of CACTA Elements

The CACTA transposons, so named for a highly conserved motif at element ends, comprise one of the most abundant superfamilies of Class 2 (cut-and-paste) plant transposons. CACTA transposons characteristically include subterminal sequences of several hundred nucleotides conta ...

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DigestionLigationAmplification (DLA): A Simple Genome Walking Method to Amplify Unknown Sequences Flanking Mutator (Mu) Transposons and Thereby Facili

Digestion–ligation–amplification (DLA), a novel PCR-based genome walking method, was developed to amplify unknown sequences flanking known sequences of interest. DLA specifically overcomes the problems associated with amplifying genomic sequences flanking high copy n ...

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Genetic and Molecular Analyses of UniformMu Transposon Insertion Lines

The UniformMu transposon population is a large public resource for reverse genetics and functional genomics of maize. Users access the collection of UniformMu genetic stocks that are freely distributed by the Maize Cooperation Stock Center using online tools maintained at MaizeGDB ...

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Using MuDR/Mu Transposons in Directed Tagging Strategies

An introduction to MuDR/Mu transposons as mutagens is provided along with protocols for using these elements to tag maize genes. Selection for retention of Mutator activity is described as well as details for establishing and screening tagging populations efficiently.

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Comparative Gel-Based Phosphoproteomics in Response to Signaling Molecules

The gel-based proteomics approach is a valuable technique for studying the characteristics of proteins. This technique has diverse applications ranging from analysis of a single protein to the study of the total cellular proteins. Further, protein quality and to some extent distribut ...

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Structural and Functional Characterization of Receptor Kinases with Nucleotide Cyclase Activity

There has been an increase in the identification and characterization of plant receptor kinases possessing nucleotide cyclase activity. This has necessitated the development of robust methodologies for the structural and functional characterization of this biologically ...

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An Affinity Pull-Down Approach to Identify the Plant Cyclic Nucleotide Interactome

Cyclic nucleotides (CNs) are intracellular second messengers that play an important role in mediating physiological responses to environmental and developmental signals, in species ranging from bacteria to humans. In response to these signals, CNs are synthesized by nucleotid ...

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Computational Identification of Candidate Nucleotide Cyclases in Higher Plants

In higher plants guanylyl cyclases (GCs) and adenylyl cyclases (ACs) cannot be identified using BLAST homology searches based on annotated cyclic nucleotide cyclases (CNCs) of prokaryotes, lower eukaryotes, or animals. The reason is that CNCs are often part of complex multifunctional ...

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Identification of Cyclic Nucleotide Gated Channels Using Regular Expressions

Cyclic nucleotide-gated channels (CNGCs) are nonselective cation channels found in plants, animals, and some bacteria. They have a six-transmembrane/one-pore structure, a cytosolic cyclic nucleotide-binding domain, and a cytosolic calmodulin-binding domain. Despite th ...

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Identification and Characterization of Cyclic Nucleotide Phosphodiesterases

Cyclic nucleotide phosphodiesterases regulate cellular levels of small molecule second messengers that control important biological processes in all kingdoms of life. Identifying and characterizing these enzymes is necessary for basic research and pharmaceutical app ...

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Inferring Biological Functions of Guanylyl Cyclases with Computational Methods

A number of studies have shown that functionally related genes are often co-expressed and that computational based co-expression analysis can be used to accurately identify functional relationships between genes and by inference, their encoded proteins. Here we describe how a compu ...

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Detection of Reactive Oxygen Species Downstream of Cyclic Nucleotide Signals in Plants

Cyclic nucleotides act in plant cell signal transduction cascades by activating cyclic nucleotide gated cation-conducting ion channels (CNGCs). Activation of CNGCs results in inward cation (including Ca2+) conductance across the plasma membrane. Elevation of cytosolic Ca2+ is ...

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Measurement of Nitric Oxide in Plant Tissue Using Difluorofluorescein and Oxyhemoglobin

Nitric oxide (NO) is now well established as a signalling molecule in plants, regulating various physiological processes ranging from development to responses to pathogens and changes in the physical environment. Various methods for the detection of NO in plant tissue have been descri ...

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Infrared Gas Analysis Technique for the Study of the Regulation of Photosynthetic Responses

Homeostatic maintenance of physiological and biochemical processes is a key requirement for survival and adaptive responses of multicellular organisms such as plants. These important processes are in part mediated by various plant enzymes and hormones, many of which are in part, con ...

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