微生物学技术

Poxvirus expression vectors have gained widespread use for expression of foreign proteins and as delivery vehicles for vaccine antigens. We have developed a novel method using the poxvirus as a library vector for functional selection of specific cDNA. Poxviruses have several unique and ...

Recombinant DNA technology has made it possible to develop molecular cloning vectors that allow the expression of heterologous genes in a variety of animal viruses. This chapter discusses the use of vaccinia virus encoding bacteriophage T7 RNA polymerase as an expression vector system. A ...

Vaccinia virus (VV) has proven to be a very useful tool for the expression and analysis of foreign gene products. The most common method used to produce recombinant viruses involves the insertion of foreign genes into the thymidine kinase (TK) gene of the VV via homologous recombination. This is acc ...

The standard approach for the isolation of vaccinia virus recombinants involves homologous recombination between a transfected plasmid and the replicating viral DNA. In a typical infection/transfection experiment, recombinant viruses only account for a tiny proportion (10 ...

Vaccinia virus, the prototype Orthopoxvirus, is widely used in the laboratory as a model system to study various aspects of viral biology, virus-host interactions, and as a protein expression system and a vaccine vector. The ubiquitous use of vaccinia viruses in the laboratory raises certain ...

Human infection has been reported with groups A, B, and C rotaviruses (RVs). Of these, Group A RVs are the most important, being a major cause of severe gastroenteritis (GE). Each year, Group A RVs are estimated to cause approx 870,000 deaths worldwide in children less than 5 years (yr) of age, mostly in developi ...

Rotaviruses (RVs) are the chief etiologic agent of viral gastroenteritis in infants and young children, and in the young of a large variety of animal species. Since the discovery of RVs in man 25 yr ago, much has been learned about their genome and protein composition; their three-dimensional struc ...

Since the discovery of animal rotaviruses (RVs) in the 1960s (1,2), and of human rotaviruses (HRVs) 25 yr ago (3,4), much has been learned about virus structure, classification, evolution, replication, pathogenesis, and specific immune responses, and their correlation with protection and ...

Rotavirus (RV), a double-stranded (ds)RNA virus in the family Reoviridae, is a complex, relatively large (diameter, including spikes = 1000 �), nonenveloped icosahedral virus. Once RV was recognized as a major human pathogen, it was extensively studied using modern molecular genetic and b ...

The effort to understand the molecular biology of rotaviruses (RVs) has led to the development of procedures that can be used to study the replication and transcription of the RV genome, the assembly and structure of the rotavirion, and the structure and function of RV proteins. Because it is not poss ...

Rotavirus (RV) is a triple-protein-layered icosahedral virus, for which studies have established that the two outer-layer proteins, viral protein 4 (VP4) and viral protein 7 (VP7), are required for viral infectivity (1,2). VP7, a glycoprotein, is the major component of the outer-layer, but its r ...

Mixed infection of tissue culture cells is the primary means of studying genetic and nongenetic interactions between viral mutants. The purpose of the mixed infection is to place two different viral genomes into the same cell, where interactions between the genomes and their encoded gene pr ...

Because of the limitations in studying human rotavirus (HRV) pathogenesis and mucosal immunity in the natural host (infants and children), various animal models have been utilized to investigate rotavirus (RV) disease pathogenesis and immunity. Mice and rabbits serve as useful mode ...

Studies of natural rotavirus (RV) infection in children have shown that protection against subsequent RV disease occurs (1). Assessment of humoral immune responses has included study of the importance of circulating vs intestinal antibodies (Abs), serotype-specific vs group-sp ...

Rotaviruses (RVs) are important human pathogens. The murine model of RV infection has been very useful in clarifying the mechanisms that mediate clearance of primary RV infection, and the mechanisms that mediate immunity to reinfection. The use of immunodeficient strains of mice, immun ...

The high morbidity and mortality of rotavirus (RV) infections has spurred the development of RV vaccines (1–13). Although children naturally infected with RV commonly undergo multiple infections, primary infections in children generally induce disease, and children are normally ...

Schistosoma mansoni is a trematode parasite with a freshwater snail intermediate host but whose definitive host is human. Schistosomiasis is a chronically debilitating, and often fatal disease affecting 200- 300 million people in many of the developing countries. In order to carry out la ...

When studies on the biochemistry, molecular biology, and genet- ics of the blood forms of malaria parasites are planned, two features of their biology should be remembered. These forms consist of parasites in a continuous cycle of development, from merozoites through ring-stages, and troph ...

Examples of already-characterized parasite T-cell epitopes (1–5; see also refs. in 6) fit with current models of antigen recognition, in which T-cells recognize foreign proteins only in the form of peptides associated with major histocompatibility complex (MHC) antigens. Peptides bo ...

This chapter aims to provide a step-by-step procedure in preparing protein samples of limited quantities for partial sequence analysis. Its content will cover a brief introduction on various types of sequenators thus far developed, with the emphasis on the latest technology of gas-liqu ...