实验基础

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 ...

The transcriptome has shown tremendous potential for the comprehensive investigation of gene expression profiles and transcriptional levels in comparative biology, the identification of regulatory mechanism of transcriptional regulators, and the evaluation of target ...

The continuous emergence of multidrug-resistant bacterial pathogens is a major problem in public health. Many mechanisms may be involved in such resistance in Staphylococcus aureus. Increasing data have shown that S. aureus can invade different types of host cells, which may contribu ...

Here, we describe a solid-phase microextraction-gas chromatography/mass spectrometry (SPME-GC/MS) analytical approach that identifies and analyzes volatile compounds in the headspace above a live fungal culture. This approach is a sensitive, solvent-free, robust techni ...

Ergot alkaloids are indole derivatives with diverse structures and biological activities. This chapter describes the procedure from fungal cultivation to purified ergot alkaloids, as exemplified by fumigaclavine A in Penicillium commune. Furthermore, useful notes for work ...

Non-ribosomal peptide synthetases (NRPSs) are important enzymes of production machinery for natural products including clinically used antibiotics, antifungal, and anticancer agents. NRPS products are usually further modified by tailoring enzymes, resulting in the form ...

Filamentous fungi have long been recognized to be a rich source of secondary metabolites with potential medicinal applications. The recent genomic sequencing of several Aspergillus species has revealed that many secondary metabolite gene clusters are apparently silent under s ...

The FERMEX (Fermentation Extract) program was a highly successful source of microbial natural product molecules for pharmaceutical lead discovery. The program was based on the observation that solid fermentations of fungi generally exhibited more complex metabolite profiles ...