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丁香实验推荐阅读
Strategies for the Isolation and Characterization of Antimicrobial Peptides of Invertebrates

Resistance of bacteria to antibiotics has become one of the main problems in human health. In addition, in agronomy, microbial diseases are largely responsible for the decrease in agricultural production. The discovery of new antibiotic families is a way to circumvent such problems and ant ...

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HPLC Methods for Purification of Antimicrobial Peptides

The advent of high performance liquid chromatography (HPLC) has greatly accelerated the discovery, purification, and characterization of antimicrobial peptides. Virtually every modern study of an antimicrobial peptide includes or was preceded by a description of its purific ...

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Origins and Development of Peptide Antibiotic Research: From Extracts to Abstracts to Contracts

Peptide antibiotic research, which in the larger sense includes protein antibiotic research, actually began during the late 19th century with the work of Ehrlich, Metchnikov, Kanthack, and Petterson. Now it has been absorbed into the fields of microbiology, immunology, histochemist ...

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Neutralization of the In Vivo Activity of E. Coli-Derived Lipopolysaccharide by Cationic Peptides

It has been established that at least three of the neutrophil-derived cationic antibiotic proteins, in addition to their antibiotic activity, can bind lipopolysaccharide. It is generally accepted that the binding to lipopolysaccharide, an outer membrane component of Gram-negat ...

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Assay Systems for Measurement of Chemotactic Activity

The traditional view of the polymorphonuclear leukocyte (PMN) or neutrophil has been a cell whose primary function was to ingest and kill bacteria. It is becoming increasingly clear that a number of these granule constituents that were originally studied because of their antibiotic acti ...

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Assay of Antibacterial Activities of the Bactericidal/Permeability-Increasing Protein in Natural Biological Fluids

All living organisms are constantly confronted by invading microbes. In mammals, host defense against microbial invasion requires specialized cellular and extracellular elements in blood and a complex signalling machinery that triggers their mobilization to extravascul ...

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The Culture and Preparation of Gametocytes of Plasmodium falciparum for Immunochemical, Molecular, and Mosquito Infectivity Studies

Since the development of methods for in vitro cultivation of the asexual blood stages of P.faZciparum (1) and the production of mature gametocytes from such cultures capable of infecting mosquitoes (2), it has been possible to conduct a wide range of studies on the sexual stages of P. filciparum. The ...

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Charting Methods for Internal Quality Control for Competition ELISA

This chapter deals with relatively simple ways to use control charts to monitor the performance of ELISAs. A rinderpest competition ELISA, for the estimation of antibodies in serum samples, is used to demonstrate the methods. This assay is available in a kit form. Constant evaluation of the use of t ...

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Practical Exercises

The aim of this chapter is to illustrate the principles of ELISA by (1) showing worked examples of each assay, including diagrams of plates and representational data from assays, (2) analyzing such data in terms of important rules that are learned at each stage, and (3) providing full working instruc ...

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Titration of Reagents

This chapter examines in detail the necessary practical skills needed to facilitate the development and sustainability of ELISAs. Basic methodologies to set up all systems are discussed so that the reader can both investigate the possibilities of tests using his or her reagents and use with ...

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Systems in ELISA

This chapter defines the terms and examines the configurations used for most applications of ELISA. Such a chapter is important because the possibilities inherent in the systems of ELISA must be understood in order to maximize their versatility in assay design. All heterogeneous systems h ...

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Test Questions

This chapter consists of a test concerning ELISA and the sciences needed to perform assays. The test can be used to gage knowledge on ELISA. Suggested points are given for the benefit of educators. Of course, the test can be adapted. All the answers can be obtained by reading this book. It may be useful to test ones ...

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Pseudotyped Vesicular Stomatitis Virus for Analysis of Virus Entry Mediated by SARS Coronavirus Spike Proteins

Severe acute respiratory syndrome (SARS) coronavirus (CoV) contains a spike (S) protein that binds to a receptor molecule (angiotensin-converting enzyme 2; ACE2), induces membrane fusion, and serves as a neutralizing epitope. To study the functions of the S protein, we describe here the gen ...

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Identification of Sugar Residues Involved in the Binding of TGEV to Porcine Brush Border Membranes

Coronaviruses most often infect the respiratory or intestinal tract. Transmissible gastroenteritis virus (TGEV), a group 1 coronavirus, infects the porcine small intestine. Piglets up to the age of 3 weeks die from diarrhea caused by the viral gastroenteritis unless they are protected ...

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Systematic Assembly and Genetic Manipulation of the Mouse Hepatitis Virus A59 Genome

We have developed a DNA assembly platform that utilizes the nonspecific, highly variable sequence signatures of type IIs restriction enzymes to assemble a full-length molecular clone of murine hepatitis coronavirus (MHV) strain A59. The approach also allows changes to be engineered i ...

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Engineering Infectious cDNAs of Coronavirus as Bacterial Artificial Chromosomes

The construction of coronavirus (CoV) infectious clones had been hampered by the large size of the viral genome (around 30 kb) and the instability of plasmids carrying CoV replicase sequences in Escherichia coli. Several approaches have been developed to overcome these problems. Here we de ...

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Transient Dominant Selection for the Modification and Generation of Recombinant Infectious Bronchitis Coronaviruses

We have developed a reverse genetics system for the avian coronavirus infectious bronchitis virus (IBV) in which a full-length cDNA corresponding to the IBV genome is inserted into the vaccinia virus genome under the control of a T7 promoter sequence. Vaccinia virus as a vector for the full-leng ...

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Generation of Recombinant Coronaviruses Using Vaccinia Virus as the Cloning Vector and Stable Cell Lines Containing Coronaviral Replicon RNAs

Coronavirus reverse genetic systems have become valuable tools for studying the molecular biology of coronavirus infections. They have been applied to the generation of recombinant coronaviruses, selectable replicon RNAs, and coronavirus-based vectors for heterologous g ...

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Manipulation of the Coronavirus Genome Using Targeted RNA Recombination with Interspecies Chimeric Coronaviruses

Targeted RNA recombination has proven to be a powerful tool for the genetic engineering of the coronavirus genome, particularly in its 3′ part. Here we describe procedures for the generation of recombinant and mutant mouse hepatitis virus and feline infectious peritonitis virus. Key to the t ...

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Production of Monospecific Rabbit Antisera Recognizing Nidovirus Proteins

The importance of monospecific antisera for the experimental analysis of viral proteins is undisputed. They make it possible to identify and analyze the target protein against a background of a large number of other proteins, either in whole fixed cells or in cell lysates. This chapter descri ...

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