微生物学技术

High-capacity adenoviral vectors (HC-Ad), also known as “helper-dependent” (HD-Ad), “gutless”, “gutted”, or “third-generation” Ad vectors, are devoid of all viral coding sequences and have shown promising potential for a wide variety of different applications—from classic gene th ...

The use of chimeric pseudotyped vectors is a common way to modify the adenoviral tropism by replacing the fiber protein. In this chapter the procedure to generate a chimeric adenovirus pre-stock from a plasmid containing the adenoviral genome is described. Also, the chimeric adenovirus rep ...

In this chapter we compile a battery of biophysical and imaging methods suitable to investigate adenovirus structural stability, structure, and assembly. Some are standard methods with a long history of use in virology, such as embedding and sectioning of infected cells, negative stain ...

Analysis of proteins and their posttranslational modifications is important for understanding different biological events. For analysis of viral proteomes, an optimal protocol includes production of a highly purified virus that can be investigated with a high-resolving ana ...

Adenoviral vectors hold immense potential for a wide variety of gene therapy based applications; however, their efficacy and toxicity is dictated by “off target” interactions that preclude cell specific targeting to sites of disease. A number of “off target” interactions have been desc ...

During human adenovirus type 3 (Ad3) infection, an excess of penton base and fiber proteins are produced. These form dodecahedral particles composed of 12 pentamers of penton base and 12 trimers of fiber protein. Beside this “natural” expression, the adenovirus dodecahedron can be express ...

Vectors derived from the canine adenovirus serotype 2 (CAV-2) possess a high neurotropism and efficient retrograde transport that lead to widespread neuronal transduction in the central nervous system (CNS) of various animals. These abilities are due to the engagement of virions to the c ...

The encapsidation of adenovirus (Ad) DNA into virus particles depends on cis-acting sequences located at the left end of the viral genome. Repeated DNA sequences in the packaging domain contribute to viral DNA encapsidation and several viral proteins bind to these repeats when analyzed us ...

Defining the molecular toxicity of viral vectors that are or will be in use for clinical trials is a prerequisite for their safe application in humans. DNA chips allow high-throughput evaluation of the profile of transduced cells and have contributed to underlining specific aspects of vector ...

Virotherapy in one of the main current applications of recombinant adenoviruses. Oncolytic adenovirus are designed to target tumors, replicate selectively in tumor cells, and elicit immune responses against tumor antigens. Transgene expression in replication-competent o ...

Recently, more emphasis has been given to understand molecular genetics and the contribution of a gene in the disease process. In fact, increased understanding of bacterial pathogenesis and intracellular communication has revealed many potential strategies for development of n ...

Immunofluorescence microscopy is a widely used laboratory method which allows detection and visualization of specific antigens. The method employs the specificity of antibodies to deliver fluorophore to a specific target and then visualize it with a microscope. The power of the tech ...

Antimicrobial susceptibility test methods such as disk diffusion, broth microdilution, and oxacillin screen plate require 24 h of incubation after having the organism growing in pure culture. Rapid and accurate identification of MRSA isolates is essential not only for patient care, b ...

Over the past decade, the emergence of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has changed the landscape of S. aureus infections around the globe. Initially recognized for its ability to cause disease in young and healthy individuals without ...

Methicillin-resistant Staphylococcus aureus (MRSA) is a major pathogen responsible for both hospital and community onset disease. Resistance to methicillin in S. aureus is mediated by PBP2a, a penicillin-binding protein with low affinity to beta-lactams, encoded by the mecA gene. A ...

One of the foremost challenges of drug discovery in any therapeutic area is that of solidifying the correlation between in vitro activity and clinical efficacy. Between these is the confirmation that affecting a particular target in vivo will lead to a therapeutic benefit. In antibacterial ...

Staphylococcus aureus is an important pathogen linked to serious infections both in the hospital and the community settings. The challenge to treat infections caused by S. aureus has increased because of the emergence of multidrug-resistant strains such as methicillin-resistant S. ...

This chapter reviews data on the treatment of infections caused by drug-resistant Staphylococcus aureus, particularly methicillin-resistant S. aureus (MRSA). This review covers findings reported in the English language medical literature up to January of 2013. Despite the emer ...

Metabolomics is becoming increasingly important as it provides a comprehensive analytical platform to better understand the biological functioning of a cell or organism. In recent years, microbial metabolomics has received much attention in research areas from new drug discove ...

Over the last two decades, numerous genomes of pathogenic bacteria have been fully sequenced and annotated, while others are continuously being sequenced. To date, the sequences of more than 8,500 whole bacterial genomes are publicly available for research purposes. These efforts in hig ...