疾病及处理

Animal models are invaluable tools for translational research, allowing investigators to recapitulate observed clinical scenarios within the laboratory that share attributes with human disease. Here, we describe a mouse model of post-arthroplasty Staphylococcus epider ...

The genetic manipulation of Staphylococcus epidermidis for molecular experimentation has long been an area of difficulty. Many of the traditional laboratory techniques for strain construction are laborious and hampered by poor efficiency. The ability to move chromosomal gene ...

Rapid screening of biofilm forming capacity by Staphylococcus epidermidis is possible using in vitro assays with 96-well plates. This method first developed by Christensen et al. in 1985 is fast and does not require specialized instruments. Thus, laboratories with standard microbio ...

A common in vitro method to study Staphylococcus epidermidis biofilm development is to allow the bacteria to attach and grow on a solid surface in the presence of a continuous flow of nutrients. Under these conditions, the bacteria progress through a series of developmental steps, ultimately ...

Transposon mutant libraries are valuable resources to investigators studying bacterial species, including Staphylococcus epidermidis, which are difficult to genetically manipulate. Although sequence-defined transposon mutant libraries have been constructed in ...

To perform mechanistic studies on the biology of bacteria including metabolism, physiology, and pathogenesis, it is essential to possess the tools required for genetic manipulation. Introduction of plasmid DNA into Staphylococcus epidermidis for subsequent genetic manipul ...

Isolation of RNA (ribonucleic acid) is a valuable technique to study gene regulation and functional RNAs. It is important to obtain pure samples of RNA for downstream applications, while avoiding the negative effects of ribonucleases (RNases). Here we describe several methods of extrac ...

The variable region (Fv) portion of an antibody is comprised of the antibody VH and VL domains and is the smallest antibody fragment containing a complete antigen-binding site. To stabilize the association of the recombinant VH and VL domains, they have been linked in single-chain Fv constructs ...

The problem of amplifying a specific antibody in a population of millions of other antibodies has been solved by the immune system using the process of clonal selection Binding of an antigen to an IgM receptor on the surface of B-lymphocytes stimulates the proliferation and differentiation of ...

The SFV Expression System is a DNA expression system used to produce recombinant protein in eukaryotic cells (1,2). The SFV system is based on the Semliki Forest Vnus (SFV), which has several features that provide distinct advantages for a good cDNA expression system. These are:

Baculovirus vectors are now widely used to direct the expression of heterologous genes in cultured insect cells and insect larvae. In most cases, heterologous genes placed under transcriptional control of the polyhedrm promoter of the Autographa californica nuclear polyhedrosis ...

The introduction of genetic engineering techniques has allowed the controlled and efficient production of recombinant proteins. This presents scientists with the opportunity to use a wide range of proteins for a number purposes, previously unavailable because of problems relat ...

For the last two decades several highly effictent expression systems have been developed allowing the production of various proteins for diagnostic, therapeutic, and preventive purposes. These expression systems are mostly based on bacteria as well as on various eucaryotic cells. P ...

In many cases, the analysis of a specific protein is impeded by the inability to purify large amounts of it from a native source. Proteins of interest may be present in minute quantities and/or purification may be plagued with technical problems. Recombinant DNA methodologies have enabled rese ...

The polymerase chain reaction (PCR) is used to amplify a segment of DNA that lies between two regions of known sequence (1–3). It requires two oligonucleotide primers that flank the DNA fragment to be amplified and employs repeated cycles of heat denaturation of the DNA, annealing of the primers to th ...

This chapter focuses on methods for epitope mapping on novel viral polypeptrdes and on fine tuning sensitivity and spectficity of the identified peptide antigens for application in virus diagnosis. Because of the development of efficient methods of multiple peptrde synthesis (1–3), a ...

The polymerase chain reaction (PCR), originally introduced by Satki et al. (1) and subsequently automated by Mullis and Faloona (2), has emerged as a powerful tool in molecular genetics for the exponential in vitro amplification of specific sequences of Interest from minute quantrties of DNA ...

Accurate and early diagnosis of a disease state such as a viral infection, or in a more complicated situation cancer, means live saving because proper medical interventions can be applied in a timely manner before it is too late to treat the disease. Thus it is crucial that good diagnostic markers for any ...

At present the protein expression systems used commonly by researchers incorporate an affinity tail fused to the protein of interest. These affinity tails provide a convenient and efficient method for the purification of the expressed fusion protein using affinity chromatography. ...

An immunoassay may be defined as an assay that employs an immunological reagent, usually an antibody, to confer specificity for the ligand being measured. As a corollary to this, the discovery, and subsequent development, of monoclonal antibodies (MAbs) has greatly expanded the applicat ...