生物化学技术

The identification of differentially expressed genes is a key step in the understanding of specific molecular mechanisms. Various methods have been developed to search for differences in expression but most of them are time or money consuming. We present here an alternative technique th ...

High levels of foreign gene expression in mouse hepatocytes can be achieved by “hydrodynamics-based transfection,” the rapid injection of a large volume of a naked deoxyribonucleic acid (DNA) solution into the tail vein. Rats are more tolerant of the frequent phlebotomies required for mo ...

This work describes a procedure for the generation of site-specific mutations into the chromosome of Streptococcus pneumoniae that does not involve the use of an antibiotic resistance marker. A linear fragment of transforming deoxyribonucleic acid (DNA) is constructed by polymer ...

Dendritic cell-based vaccines are being evaluated in clinical trials to determine their ability to activate clinically relevant tumor antigen-specific immune responses. Although some groups isolate dendritic cells from peripheral blood, most have found it more efficient to ge ...

Bacterial superantigens (SAGs) bind to cognate Vβ elements of T-cell receptors on T-cells and to major histocompatibility complex (MHC) class II molecules on antigen-presenting cells to activate T-cell subsets expressing the Vβ elements. We examined the possibility that the direct b ...

Electroporation is a method for introducing DNA into cells by using a high-voltage electric field. This method is very simple and easily manipulated. We describe here a method for the modification of tumor cells with the Fas/Apo-1 (CD95) antigen-gene and Fas ligand (FasL)-gene transfection t ...

The knowledge of tumor-associated T-cell epitopes is important for the understanding of tumor biology and the development of cancer vaccines. We describe here a biochemical approach for the identification of tumor-associated T-cell epitopes. Peptides are extracted from immunoa ...

Cytotoxic T-cells are the most important effector cells in immune responses against tumors. The identification of tumor-associated epitopes for these cells, therefore, has become a key aspect of the development of cancer vaccines. Here, we describe a new approach to the determination of t ...

Metastatic mammary tumors in rats were treated by laser immunotherapy. Tumors injected by a laser-absorbing dye and an immunoadjuvant were irradiated non-invasively by a near-infrared laser. The successfully cured rats developed a long-term resistance to repeated tumor challen ...

Immune stimulation is a promising prospect in cancer therapy. Immunotherapy may be local or systemic, aspecific or targeted and may use monoclonal antibodies or vaccines. The aim of using vaccines is to stimulate the body to produce its own antibodies. Autologous tumor-cell vaccination h ...

Multilocus sequence typing (MLST) is used by the Scottish Meningococcus and Pneumococcus Reference Laboratory (SMPRL) as a routine method for the characterization of certain bacterial pathogens. The SMPRL recently started performing MLST on strains of Streptococcus pneumon ...

Pyrosequencing™ technology is a powerful genotyping tool that requires the generation of single-stranded DNA. Currently, two simple, solid-phase-based methods are available for this, but they require special equipment, they are not automated, and they are relatively expensive be ...

The ability to identify disease-associated genes using microarray technology is dependent on isolating high-quality total RNA from the diseased tissue under study. However, it is not always possible to obtain large amounts of affected tissue from patients and therefore the quality and ...

Epitope tagging is a valuable tool for quick detection, isolation, and analysis of protein-protein interaction, without prior knowledge of the target protein. The FLAG epitope tag, one of the most widely used tags, is an eight amino acid peptide that can be detected by anti-FLAG monoclonal antib ...

The mapping of genetic loci within organisms has been accelerated by the advent of Radiation Hybrid (RH) panels. These panels are available for humans and non-humans including mice, baboon, rat, and canine. This article contains a general protocol for the use of the Genebridge 4 whole genome RH pan ...

A simple and efficient method is described for isolating high quality RNA from bilberry fruit. The procedure is based on the use of hexadecyltrimethyl ammonium bromide (CTAB), polyvinylpyrrolidone (PVP), and β-mercaptoethanol in an extraction buffer in order to eliminate the polysac ...

The isolation of drug-resistant strains of herpesviruses, including Herpes Simplex Virus type 1 (HSV-1) and type 2 (HSV-2), Varicella-Zoster Virus (VZV), and cytomegalovirus (CMV), has been reported with increasing frequency in immunocompromised patients and is a matter of major conc ...

Chemokines (e.g., IL-8) were originally identified as chemotactic proteins obtained from various different natural sources. Today, using the genome walking strategy an ever-increasing number of novel genes encoding chemokines have been discovered that were expressed in bacter ...

Microsatellites are highly polymorphic repetitive DNA segments dispersed throughout the genome and have been widely used for genetic linkage analysis and allele loss. Instability of microsatellites sequences has been linked to deficiencies in DNA mismatch repair, and is observ ...

A rapid method for the extraction and purification of DNA from environmental samples for molecular cloning applications was developed. The indigenous cells from plant debris, organic materials, sediments, and soils were lysed directly by using DAS-IZ solution and the nucleic acids we ...