植物学技术

In this chapter we present a detailed protocol for the large-scale identification of proteins present in rice leaf and root tissue samples using 2D liquid chromatography-tandem mass spectrometry of protein extracts. This is performed using biphasic (strong cation exchange/rever ...

Tandem mass spectrometry is an efficient technique for the identification of peptides on the basis of their fragmentation pattern (MS/MS scan). It can generate individual spectra for each peptide, thereby creating a powerful tool for protein identification on the basis of peptide chara ...

We describe a procedure allowing extraction of total proteins that performs efficiently with a large variety of plant tissues, based on simultaneous precipitation and denaturation with TCA and 2ME in cold acetone. We also describe protein solubilization prior to IEF, either in classic ...

The Western blotting/sequencing technique using polyvinylidene difluoride (PVDF) membrane is one of the most popular technique for Edman sequencing. A protein sample is transferred from a 2D polyacrylamide gel electrophoresis (2D-PAGE) gel onto a PVDF membrane by electroblott ...

We present the background for multivariate data analysis on proteomics data with a hands-on section on how to transfer data between different software packages. The techniques can also be used for other biological and biochemical problems in which structures have to be found in a large amount of ...

The explosion of genomics data has led to a significant knowledge gap, with thousands of genes identified having no known function. The following chapter describes the available forward and reverse genetics strategies, which can assist researchers in assigning functions to novel gen ...

Transient expression in plants is a valuable tool for many aspects of functional genomics and promoter testing. It can be used both to over-express and to silence candidate genes. It is also scaleable and provides a viable alternative to microbial fermentation and animal cell culture for the pr ...

This chapter provides an overview of the main steps in the process to produce stably transformed plants. Most transformation methods use tissue culture to recover adult plants from regenerable explants and can be divided into three stages: (1) choice and preparation of explant tissue, (2) de ...

The BASC system provides tools for integrated mining and browsing of genetic, genomic and phenotypic data. The BASC demonstration server provides access to raw and analysed information for Brassica species and comparative information with Arabidopsis. We can use the tools within the B ...

Comparative genomics is an emerging and powerful approach to achieve crop improvement. Using comparative genomics, information from model plant species can accelerate the discovery of genes responsible for disease and pest resistance, tolerance to plant stresses such as drought, ...

To identify the genes and gene functions that underlie key aspects of legume biology, researchers have selected the cool season legume Medicago truncatula as a model system for legume research. The mission of the M. truncatula Consortium is to promote unrestricted sharing of data and inform ...

Serial analysis of gene expression (SAGE) is a powerful technique which yields a digital measure of gene expression through the sequencing of libraries of specific mRNA-derived fragments, namely SAGE tags. This chapter introduces the methods and software tools that are available for re ...

Massively parallel signature sequencing (MPSS) is a technology capable of sequencing simultaneously almost all the DNA molecules in a sample. This technology is well suited for deep profiling of mRNA and small RNA by the sequencing of cDNA tags. A series of mRNA MPSS databases has been created fr ...

Nucleotide and protein sequence databases are major resources for biological and medical research. This chapter introduces the European Molecular Biology Laboratory (EMBL) Nucleotide Sequence Database, a comprehensive primary data archive for nucleic acid sequences, and G ...

Kyoto Encyclopedia of Genes and Genomes (KEGG) is a bioinformatics resource for understanding biological function from a genomic perspective. It is a multispecies, integrated resource consisting of genomic, chemical, and network information, with cross-references to numero ...

Metabolomics is the large-scale analysis of metabolites and as such requires bioinformatics tools for data analysis, visualization, and integration. This chapter describes the basic composition of chromatographically coupled mass spectrometry (MS) data sets used in metabo ...

Molecular genetic markers represent one of the most powerful tools for the analysis of genomes. Molecular marker technology has developed rapidly over the last decade, and two forms of sequence-based markers, simple sequence repeats (SSRs), also known as microsatellites, and single nu ...

Passport and phenotypic data on germplasm and breeding lines are available from worldwide sources in various electronic formats. These data can be collated into a single database format to enable strategic interrogation to make the best use of data for effective germplasm use and enhance ...

The Gene Ontology (GO) is an established dynamic and structured vocabulary that has been successfully used in gene and protein annotation. Designed by biologists to improve data integration, GO attempts to replace the multiple nomenclatures used by specialised and large biological k ...

The accurate identification of exons and introns that comprise a complete plant gene structure can be a time-consuming and challenging task. Novel Web-based tools facilitate the process by providing a convenient interface to current transcript evidence, and portals to relevant bioi ...