Epidemiological studies, which address the role of human papillomavirus (HPV) in the pathogenesis of (pre)malignant cutaneous lesions, focus on the HPV B1 subgroup comprising the so-called epidermodysplasia verruciformis (EV)-associated HPV types. To detect and type HPV DNA in hu ...
The productive phase of human papillomavirus (HPV) infection is dependent on squamous differentiation of epithelial keratinocytes. Organotypic culture systems of primary human keratinocytes (PHKs) or immortalized keratinocytes that contain HPV genomes were developed ...
The organotypic (raft) culture system has been used to develop an in vitro system that is capable of reproducing the entire human papillomavirus (HPV) life cycle, including virion morphogenesis. This system utilizes HPV-containing cell lines that are either derived from biopsies or cre ...
The study of human papillomaviruses (HPVs) has been challenging due to the differentiation-dependent aspects of their productive life cycles. The use of HPV virions, isolated from tissues, to study viral pathogenesis has been complicated due to the low numbers of HPV virions synthesized ...
The papillomavirus life cycle is tied to the differentiation of the stratified squamous epithelium that this virus infects. The ability to study the papillomavirus life cycle is facilitated by organotypic culturing techniques that allow one to closely recapitulate this terminal d ...
The generation of cell strains and established cell lines from human papillomavirus (HPV)-infected cervical biopsies and ano-genital warts is best achieved by the application of conventional protocols for keratinocyte cell culture. The optimal protocol that permits growth at cl ...
The folding of a protein to its final native state involves a series of complex steps of intra- and intermolecular interactions between the nascent polypeptide chain, its solvent environment, and the quality control machinery of the cell (1). These steps are often halting, as the unfolded prote ...
Protein folding in the viscous and crowded environment of the cell is very different from in vitro processes in which a single protein is allowed to refold at low concentration in an optimized buffer. Although Anfinsen's observation that all the information necessary for a protein to reach a prop ...
Detailed biophysical, biochemical, and structural studies rely on the preparation of milligram amounts of pure recombinant proteins. Many useful overexpression systems have been developed for this purpose (1–3), and of these, bacterial overexpression is still the most convenie ...
Methods for the efficient overexpression and purification of recombinant proteins are of paramount importance for biotechnology. In particular, for the era of functional genomics that we have entered after sequencing complete genomes, this has become a routine matter. High-thro ...
Affinity chromatography, the method of purifying target proteins from complex mixtures using immobilized affinity ligands on chromatographic supports, is perhaps the most common of all affinity techniques (1–3). Many affinity chromatography systems are comprised of activa ...
The use of λ repressor fusions to study protein-protein interactions in E. coli was first described by Hu and others (1). Since then, the repressor system has been employed by several laboratories to screen genomic (2–5) and cDNA libraries (6) for homotypic or heterotypic interactions. λ repress ...
Though widely used, expression of heterologous genes in E. coli can be cumbersome and often fails to yield significant amounts of the desired protein. There are a variety of reasons why a particular heterologous gene fails to yield significant amount of protein, including susceptibility of t ...
Two-hybrid systems are powerful genetic assays that allow the interaction between two proteins to be detected in vivo. Although originally described in yeast (1,2), several bacterial two-hybrid systems have recently been developed (reviewed in 3–6). This chapter will describe the use of ...
The completion of the human genome project has ushered in a new era of life science (1,2) in which the new challenge is to understand functions of the entire collection of the gene products, or the proteome. One important feature of biological research in this post-genomics era is the emphasis on unders ...
Advances and opportunities in drug discovery and functional genomics have put methods for generating molecular diversity at a premium. Both chemical and biological approaches for the production of compound libraries have been pursued. Combinatorial chemistry has been used to syn ...
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 ...
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