生物信息学技术

Obtaining well-diffracting crystals remains a major challenge in protein structure research. In this chapter, we review currently available computational methods to estimate the crystallization potential of a protein, to optimize amino acid sequences toward improved cryst ...

Nucleotide and protein sequences are the foundation for all bioinformatics tools and resources. Researchers can analyze these sequences to discover genes or predict the function of their products. The INSD (International Nucleotide Sequence Database—DDBJ/EMBL/GenBank) is an ...

This chapter reviews the methodologies for RNA structure determination by liquid-state nuclear magnetic resonance (NMR). The routine production of milligram quantities of isotopically labeled RNA remains critical to the success of NMR-based structure studies. The standard m ...

X-ray biocrystallography is the most powerful method to obtain a macromolecular structure. The improvement of computational technologies in recent years and the development of new and powerful computer programs together with the enormous increment in the number of protein struct ...

Microarrays have become a widely used technology in molecular biology research. One of their main uses is to measure gene expression. Compared to older expression measuring assays such as Northern blotting, analyzing gene expression data from microarrays is inherently more complex d ...

Hepatocyte growth factor (HGF) has mitogenic, motogenic, and morphogenic biological activities as well as helps in regenerating various tissues. In cardiovascular organs, HGF was reported to have anti-apoptotic, anti-fibrotic, and vasodilating effects. HGF has close relation ...

Electroporation is a powerful method for gene delivery to dystrophic muscle in the mdx mouse model of Duchenne muscular dystrophy. Successful transfer of reporter and therapeutic plasmids and antisense oligonucleotides has been demonstrated. However, the efficiency falls with ...

Defibrillation shocks are commonly used to terminate life-threatening arrhythmias. According to the excitation theory of defibrillation, such shocks are aimed at depolarizing the membranes of most cardiac cells, resulting in resynchronization of electrical activity in the h ...

Studies described in the recent literature support the idea that gene therapy can lead to genuine clinical benefits when mediated by plasmid delivery in conjunction with electroporation. Plasmid-mediated muscle-targeted gene transfer offers the potential of a cost-effective p ...

DNA-based cancer vaccines have been used successfully in mice to induce cytotoxic T lymphocytes (CTLs) specific for prostate antigens. Translation of a prostate-specific antigen (PSA) DNA vaccine into a phase I clinical trial demonstrated that PSA-specific immune responses could be ...

Electroporation has been shown to be an effective method to improve the efficiency of gene expression and the immunogenicity of DNA vaccines. To optimize the procedure and test for its efficacy in more clinically relevant large animal models, we studied the effects of electroporation-me ...

Several studies of DNA vaccination against HER2/neu showed the effectiveness of immunization protocols in models of transplantable or spontaneous tumors. The DNA delivery system plays a crucial role in the success of DNA vaccination. In particular, our studies of DNA vaccination agai ...

We are presently aware of two early-phase DNA vaccine clinical trials in humans using electroporation-enhanced vaccine delivery. Moreover, two phase I immunogenetherapy studies are in progress and several tolerability studies have been performed on healthy volunteers. We have u ...

DNA immunization with in vivo electroporation is an efficient alternative protocol for the production of monoclonal antibodies (mAb). Generation of mAb by DNA immunization is a novel approach to circumvent the following technical hurdles associated with problematic antigens: l ...

In bioaffinity immobilization, the enzyme/protein is immobilized via bioaffinity interactions. A large number of affinity pairs such as lectin-sugar, antigen-antibody, and biotin-avidin are known. The use of affinity tags to create fusion proteins that can bind to the desired matrix ...

In this chapter, the combined use of the selectivity of metal chelate affinity chromatography with the capacity of epoxy supports to immobilize poly-His-tagged proteins via multipoint covalent attachment is shown. This has been achieved by designing tailor-made chelate-epoxy sup ...

Subunit dissociation of multimeric proteins is one of the most important causes of inactivation of proteins having quarternary structure, making these proteins very unstable under diluted conditions. A sequential two-step protocol for the stabilization of this protein is propo ...

Immobilization of lipases on hydrophobic supports at low ionic strength permits one- step purification, immobilization, hyperactivation, and stabilization of most lipases. This selective adsorption occurs because the hydrophobic surface of the supports is able to promote the ...

The prospects of a new commercially available support (amino-epoxy-Sepabeads� ) for enzyme immobilization are discussed in this chapter. These supports have a layer of epoxy groups over a layer of ethylenediamine that is covalently bound to the support. Thus, the support has a high anionic ex ...

Chemical modification and immobilization of proteins have been usually utilized as parallel techniques to improve enzyme stability. In this chapter, we show that chemical modification of the protein surface to greatly increase its reactivity with the groups of a support activated wi ...