植物生理

Ribosome-inactivating proteins (RIPs) are cytotoxic N-glycosidases identified in plants, fungi, and bacteria. RIPs inhibit protein synthesis by virtue of their enzymatic activity, selectively cleaving a specific adenine residue from a highly conserved, surface-exposed, s ...

A protocol for the establishment of in vitro shoot cultures of Catharanthus roseus is described. Shoots can be maintained for more than 1 yr without evidence of tissue vitrification, disaggregation, or callus formation. Vindoline was the main alkaloid accumulated, reaching values sim ...

Identifying parasitism genes encoding proteins secreted from a plant-parasitic nematode’s esophageal gland cells and injected through its stylet into plant tissue is the key to understanding the molecular basis of nematode parasitism of plants. Parasitism genes have been cloned ...

Inactivation of a chromosomal gene is a useful approach to study the function of the gene in question and can be used to produce a desired phenotype in the organism. This chapter describes how to generate such mutants of the cyanobacterium Synechococcus sp. PCC 7002 and the green sulfur bacterium Chl ...

A method to prepare photosystem I (PSI) particles is described. Spinach leaves are used to prepare broken chloroplasts that are then solubilized by using a detergent (Triton X-100). Solubilized chloroplasts are then applied on an ion-exchange column. Eluted by a linear concentration gra ...

The available procedures for isolation and purification of photosystem I (PSI) from Chlamydomonas reinhardtii are time consuming and usually require several hours of sucrose gradient ultracentrifugation steps. This may lead to structural and functional impairment, includ ...

This chapter contains the description of several methods used for the isolation of functional photosystem (PS)II core particles from wild-type (wt), PSI-less, and CP47 histidine-tagged cells of the cyanobacterium Synechocystis sp. PCC 6803. These protocols discuss the cultivati ...

Methods for the isolation of highly active oxygen-evolving photosystem (PS)II membranes from higher plants and the purification of the oxygen-evolving complex (OEC) subunits are described. Membrane samples used as the material for various in vitro studies of PSII are prepared by solu ...

This chapter deals with the preparation of photochemically active reaction center particles (Type III) that basically contain only two subunits, PsaA and PsaB. The preparation is obtained by further treating the Type II preparation described in the preceding chapter with a chaotropic ...

Methods to prepare photosystem (PS)I reaction center (RC) subunit polypeptides are described. For PsaC, PsaD, PsaE, and PsaY, the filtrate resulting from the ultrafiltration in Chapter 6 is used for preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PA ...

Methods to isolate and purify 6- and 5-chlorophyll (Chl) D1/D2/Cyt b 559 photosystem (PS) II reaction center complexes from plants are presented, and the advantages and disadvantages of each procedure are discussed. One of the simpler 6-Chl procedures and a procedure for isolating 5-Chl compl ...

Many vaccine antigens and biopharmaceutical proteins have been expressed at high levels via the chloroplast genome and their functionality has been evaluated using in vitro assays in cell cultures (i.e., macrophage lysis assay, inhibition of vesicular stomatitis virus-induced c ...

N-glycosylation is a maturation event necessary for the correct function, efficiency, and stability of a high number of biopharmaceuticals. This chapter presented here proposes various methods to determine whether, how, and where a plant pharmaceutical is N-glycosylated. These me ...

Recombinant antibody therapeutics represent a significant success story in terms of clinical benefit delivered and revenue (profit) generated within the biopharmaceutical industry. Additionally, it is estimated that ̃30% of new drugs likely to be licensed during the next decade w ...

Over the last decade, plant-based production of pharmaceuticals has made remarkable progress as the expression of a diverse set of proteins has been demonstrated in a range of plant crops. Although the commercial exploitation is still pending, today various plant-based expression tec ...

Stable accumulation of storage proteins, lipids and carbohydrates is a hallmark of the plant seed, and is a characteristic that is typically deficient in existing platforms for recombinant protein manufacture. One of the biological sequestration mechanisms that facilitate the fo ...

We describe a general approach for the use of recombinant protease inhibitors as stabilizing agents for clinically useful proteins extracted from transgenic plant tissues. A procedure is first described to assess the overall (in)stability of heterologous proteins in transgenic p ...

Transgenic plants are gaining increasing attention from the industry as a natural bioreactor for the production of industrial and chemical products. Optimization of transgene expression in plant cells holds the key to maximizing the potential of plants for producing proteins of com ...

In this chapter, we discuss and compare the different concepts and examples as well as present the basic protocols for applying intrabody-based approaches in plants for the investigation of cell functions and plant cell–pathogen interactions. The immunomodulation strategy, a mole ...

This chapter presents a general procedure for the on-chip detection and quantitation of low-molecular-weight recombinant proteins in transgenic plant crude extracts by surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS). A prot ...