实验基础

Adenoviruses exhibit considerable intraserotypic genetic variability. Restriction enzyme analysis of the adenoviral genome is currently the most widely used procedure for the characterization of adenovirus isolates and has been extensively used for molecular epidemi ...

Adenoviruses (Ads), like other DNA tumor viruses, have evolved specific regulatory genes that facilitate virus replication by controlling the transcription of other viral genes as well as that of key cellular genes. In this regard, the E1A transcription unit contains multiple protein d ...

Here we describe a collection of methods that have been adapted to produce highly efficient nuclear and cytoplasmic extracts from adenovirus-infected HeLa cells. We describe how to produce extracts from virus-infected cells and how to analyze RNA splicing in vitro using T7 RNA polymerase ...

ELISAs offer excellent specificity and, once fully optimized, sensitivity that rivals that of bioassays. The major variables that need to be experimentally determined when developing an ELISA are the optimal number of fresh cells required per well, the optimal antigen concentrations ...

Chemokines are primarily low molecular mass proteins that are produced and usually released by a wide variety of cell types. Differential chemokine responses can be excellent early markers of immune dysfunction, allowing clinical intervention prior to expression of full blown unde ...

Immunoglobulins are a heterogeneous group of proteins. It naturally follows that the strategies for purifying them are diverse and numerous. A good knowledge of their respective physiochemical properties will obviously make the task easier. The choice between using polyclonal and ...

Specific allergen immunotherapy is an effective treatment for IgE-mediated allergic disease and involves T- and B-cell mediated events. IgE receptors on the surface of antigen-presenting cells facilitate the presentation of allergens in the presence of specific IgE antibody res ...

The genus Helicobacter currently comprises over 15 different species with members described as microaerophilic gram-negative spiral rods with a G + C content of 35–38%. The most medically important Helicobacter is Helicobacter pylori, a human pathogen that has been causally linked to t ...

Lactococcus lactis strains are the principal organisms used in cheese-making, and their performance is of vital importance to the quality of the cheese. A goal for the dairy industry has always been to improve strains of L. lactis and to stabilize beneficial traits. In recent years, genetic engin ...

The streptococci are a diverse genus of bacteria consisting of commensal and pathogenic organisms of human and veterinary origin, as well as industrially important species. Although many of the species are naturally transformable, several species do not enter a competent state. The abi ...

Diversity—the variability carried by the amino acid sequences of a synthetic antibody library—can be generated by synthetic degenerate oligonucleotides. One can experiment with different diversity designs in the variable domains of light and heavy chains (VH and VL) to generate ant ...

Antibody-based immunologic reagents are useful for identifying, isolating, or eliminating cells with particular characteristics related to different diseases. Phage display is a highly valuable technique for antibody selection related to this purpose. In brief, a diverse gro ...

Recently there has been an increased use of microarray technology as a tool to determine the presence of functional genes in a population of hard to culture communities (e.g., soil, extreme environments). A functional gene array (FGA), or GeoChip, uses probes to screen for specific functional ge ...

Many important analytes are present in the ocean water and primary examples include various marine toxins. The unique marine environment possesses an extremely high ionic strength, posing a significant analytical challenge for biosensor design. Protein-based enzymes and antib ...

There is a need for low-cost, rapid, and accurate detection of harmful organisms. Some species of the marine dinoflagellate Alexandrium produce paralytic shellfish toxins that can accumulate in shellfish tissue and can cause paralysis and death if ingested by humans. We have developed a me ...

Heavy metal contamination of oceans, lakes, and other water resources can occur by both natural and human-related processes. Human exposure to heavy metals such as mercury is known to cause a number of serious health problems. Due to its high toxicity and bioaccumulative properties, the maxim ...

The ocean contains a number of metal ions that are either beneficial or detrimental to marine lives or ecology. Developing sensors for on-site and real-time detection of these metal ions plays an important role in our understanding the ocean as well as its protection. DNAzymes, DNA molecules with ...

We present an example-based methodology for the “open-water” technique to determine net ecosystem metabolism (or net community metabolism) of coastal and oceanic environments using dissolved oxygen measurements collected with in situ sensors. We briefly discuss strategies f ...

A miniaturized and self-contained in situ genetic analyzer Integrated In Situ Analyzer-Gene (IISA-Gene) was developed to conduct PCR-based targeted sequence detection using microfluidic technology. The core element of the in situ analyzer is a microfluidic device that conducts c ...

The Monterey Bay Aquarium Research Institute’s Environmental Sample Processor (ESP) is well established as an innovative sampling and instrument platform for sensors designed for in situ monitoring of micro�organisms in the ocean. The platform can be deployed for periods up to 3 months a ...