Rapid transfer of exponentially growing E. coli cultures from physiological to low temperatures (10–15#X00B0;C) has profound consequences on cell physiology: membrane fluidity decreases, which interferes with transport and secretion, the secondary structures of nucleic ac ...

Since its invention in the early 1990s, phage display has revolutionized the generation and engineering of monoclonal antibodies (for review, see ref. 1). Without the need for laboratory animals or hybridomas, it was now possible to create antibodies binding to almost any antigen of choice. A ...

Among secondary metabolites, many interesting compounds including flavors, fragrances, and those with pharmaceutical potential can be found. Once the biosynthetic pathway of an interesting compound or group of compounds has been elucidated and the genes encoding the enzymes of the ...

The advent of combinatorial approaches to problems at the interface between chemistry and biology has had a profound impact on areas ranging from drug discovery to protein chemistry. One area of intense work has been the application of combinatorial libraries of proteins to the discovery of ...

In the near future, the nucleotide sequence of the genomes from many different organisms will be available. The next and more challenging step will be to characterize the biological role of each gene and the way in which the encoded protein functions in the cell. Dual-expression vectors for expres ...

Protein fusions are invaluable tools for the genetic studies involving the mechanisms of protein export in bacteria. In 1985, Hoffman and Wright developed an in vitro fusion approach that allowed for fusions of the gene encoding Escherichia coli alkaline phosphatase to a variety of cloned g ...

The IMPACT (Intein-Mediated Purification with an Affinity Chitin-Binding Tag) vectors are designed for the isolation of pure, functional, recombinant proteins by a single affinity chromatography step. The IMPACT technology was developed at New England Biolabs (NEB) by exploiti ...

Calmodulin is a small (148 amino acids, 17 kDa) ubiquitous protein in eukaryotes, and is considered the primary intracellular calcium sensor, making it a key regulator of intracellular signal transduction. Upon calcium binding, calmodulin can interact with a variety of proteins (1), medi ...

A major impediment to the production of recombinant proteins in Escherichia coli is their tendency to accumulate in the form of insoluble and biologically inactive aggregates known as inclusion bodies. Although it is sometimes possible to convert aggregated material into native, bio ...

Protein purification is an important tool for investigations on protein function, structure analysis, and biotechnological use. Therefore a number of different techniques have been developed for fast, reliable, and reproducible overexpression and purification of relevant ...

Escherichia coli has traditionally been a popular host for the production of heterologous proteins because of its ease of genetic manipulation and growth. Recombinant proteins produced in E. coli have been useful for a variety of purposes, including the study of protein tertiary structu ...

Fusion protein technology has been creatively applied to solve many problems encountered in the study of protein structure and function (1,2). One prevalent application is the use of fusion proteins to improve protein expression in Escherichia coli and provide convenient methods for a ...

The interactions of the hepatitis C virus (HCV) with the immune system are numerous. As one of the many results, a considerable number of autoantibodies occur in serum. This chapter describes methods to detect anti-GOR antibodies (1–3). Anti-GOR are autoantibodies that are specifically ass ...

Chimpanzees (Pan troglodytes) became established as invaluable models for the study of human viral hepatitis after it was discovered, in 1967, that the chronic hepatitis B antigen carrier state existed in a naturally infected member of this species (1–3). They were instrumental in the devel ...

HCVs constitute a genus within the Flaviviridae, with closest homology to the hepatitis G and GB viruses, and Pestiviruses. The positive-stranded RNA genome encodes at least nine proteins. Core, El, and E2 constitute the structural proteins; NS2, NS3, NS4A, NS4B, NS5A, and NS5B are nonstructur ...

Hepatitis C virus-associated end-stage liver disease is a leading diagnosis in patients undergoing liver transplantation, accounting for approx 25% of patients transplanted at major medical centers in the United States, and for 5–15% of those transplanted worldwide (1). Although HCV ...

Hepatitis C virus (HCV) possesses a single-stranded, positive-sense RNA that is 9.4 kb in length. The complete HCV genome has been cloned and sequenced and encodes for a nucleocapsid, an envelope, and five nonstructural proteins (1,2). The 5′ untranslated region of the virus is highly conserved am ...

Clinical diagnosis of HCV infection is generally accomplished by using immunoserological assays to detect the presence of anti-HCV antibodies. Such immunoserological assays have been approved for blood donor screening, thereby reducing the incidence of post-transfusion he ...

In the field of biological science, the development of new techniques (e.g., Southern blotting, molecular cloning, pulsed-field gel electrophoresis) often led to better understanding of fundamental and applied biological problems. The polymerase chain reaction (PCR) is among the t ...

The diagnosis and monitoring of hepatitis C virus (HCV) infection have been aided by the development of HCV RNA quantification assays A direct measure of viral load, HCV RNA quantification has the advantage of providing information on viral kinetics and provides unique insight into the dise ...