植物学技术

RNAi can be an effective means to regulate endogenous gene expression in maize and as such represents an important reverse genetics tool. This approach involves designing a transgenic construct that creates a double stranded RNA (dsRNA) upon transcription, and introducing the transg ...

The transgenic RescueMu lines were designed for and successfully used in our maize gene discovery project. The pBluescript-containing RescueMu transposon can be readily recovered by a procedure called plasmid rescue. Plasmid rescue is a technique for recovering bacterial plasm ...

Efficient methods to characterize transgenic plants are important to quickly understand the state of the transformant. Determining transgene copy number is an important step in transformant characterization and can differentiate between complex and simple transformati ...

This method allows high-throughput qualitative screening to identify seeds containing a transgene product in endosperm tissue. It is particularly useful for determining genetic segregation ratios or identifying seeds to be advanced in a breeding program. Tissue printing is used ...

Sensitive and reproducible genotyping tools are fundamental in interpreting and substantiating genetic data. In cases where alternative assays like PCR are not applicable, a sensitive genomic Southern protocol is needed. Our maize gene discovery work using the RescueMu transge ...

There are thousands of porphyrins and metalloporphyrins, and hundreds of new derivatives appear each year. This variety arises because the cyclic conjugated tetrapyrrole nucleus (Figure 1) can have different substituents at the eight β-pyrrole positions, at the four meso carbon ato ...

The term bilin is a collective one to describe a broad group of open chain tetrapyrroles and derives from the name “bile pigments” as the first of these compounds to be characterized were isolated from animal bile. These bilins, biliverdin (BV) and bilirubin (BR), are the sequential products of heme de ...

Heme is perhaps the most ubiquitous cofactor found in nature and the most functionally diverse. Hemoproteins are involved in cell respiration (cytochromes), oxygen-binding and transport (hemoglobin and myoglobin), oxidative bio transformations (cytochrome P-450 and perox ...

This chapter addresses basic methodology that can be used to obtain tetrapyrrole macrocycles in the porphyrin and chlorin series from natural materials and some simple methods for the total chemical synthesis of typical pyrroles and porphyrins. The aim is to provide investigators with ...

Since the 1963 seminal review of Smith and French (62), our understanding of the chlorophyll (chl) biosynthetic pathway has changed dramatically. Several factors have contributed to this phenomenon, among which are: (i) development of systems capable of Chl and thylakoid membrane bios ...

Methods of gene cloning and genetic engineering of immunoglobulins and transgenic plant techniques have given rise to new approaches in plant biotechnology (1). The high affinity and specificity of antibodies to different structures can be used to block regulation factors in plant cel ...

Plant and animal RNA viruses are increasingly used for gene delivery and expression in a variety of host cells. At a molecular level these agents can be viewed as cytosolic parasites of the ribosome, successfully competing for translation with the host messenger RNA (mRNA) population. In many vi ...

We are interested in studying plant systems as vehicles for the production of recombinant proteins of clinical relevance. There are a number of potential advantages to producing recombinant protein products in plants. Plants are more economical compared to fermentation or cell-cul ...

The discovery that the expression of a viral coat protein in transgenic plants confers protection to infection by homologous and related viruses (1–4) revolutionized the field of plant breeding. The approach of “coat protein-mediated protection” (CPMP) became an extensively studied ...

The importance of proteolytic enzymes in plant-pest and plant-pathogen interactions has recently been recognized, and control strategies based on their inhibition with protease inhibitors (PIs) have been developed or proposed to control herbivory insects (1), plant parasitic f ...

One of the most important considerations in the expression of heterologous proteins in plants is the choice of promoter. The study of promoter activity is simplified in the majority of cases by the use of a readily detectable reporter gene. Indeed, reporter genes can also be used to aid the isolation of ...

Legumes form an important element in many ecological and agricultural environments. In order to improve the range of phenotypes available for agriculture, plant breeding programs seek to produce improved varieties. Inevitably, these programs are limited by the available gene pool a ...

Electrophoresis is defined as the migration of a charged particle in an electric field. Electrophoretic techniques have been used to analyze, separate, and characterize proteins, peptides, and nucleic acids. A number of different one-dimensional electrophoresis techniques ha ...

In eukaryotic cells, secreted proteins can be subjected to numerous post-translational modifications. One of these, glycosylation, consists of the addition of sugar residues to the protein backbone, while the protein enters or travels within the secretory pathway. Glycosylation ...

Recent developments in genetic engineering and molecular biology have enabled the production of transgenic plants with improved characteristics, or cell and tissue cultures with altered or improved metabolic activity. However, no universally applicable gene transfer meth ...