基因表达差异显示实验

Several methods have been published that rely on the use of short oligonucleotide primers with arbitrary sequences to amplify discrete DNA fragments by the polymerase chain reaction (PCR) (1,2). Typically, a single arbitrary primer is used in each reaction and amplification is achieved w ...

The vectorette unit (1) consists of a pair of annealed oligonucleotides that contain two regions of complementary nucleotide sequence flanking a 29-bp noncomplementary segment (Fig. 1). The 5′ terminus of one of these complementary regions is phosphorylated and displays a restriction ...

The termini of yeast artificial chromosome (YAC) inserts can serve as probes or sequence tagged sites (STSs) that are useful in genomic analysis. For example, these elements can be used to:

In the construction and characterization of yeast artificial chromosome (YAC) contigs, it is necessary to be able to isolate and map the ends of the genomic inserts. This is important with respect to both extending contigs, and identifying chimerism. A number of techniques have now been descri ...

The detailed analysis of the DNA cloned in yeast artificial chromosomes (YACs) is performed by subcloning into vectors such as phages or cosmids, which allow a simpler purification of insert DNA in addition to allowing high resolution mapping. Cosmids or phages are still a preferred DNA source ...

The very large cloning capacity of yeast artificial chromosomes (YACs) has facilitated the analysis of complex genomes by bridging physical and genetic maps. The large size of YAC inserts also creates some unique problems, including identification of novel genes on large stretches of un ...

Gene-based in vivo electroporation has the potential to be used as a “protein-free” method to elicit immune responses and to generate monoclonal antibodies (mAb) against proteins/peptides in hosts. However, the method is very useful to raise mAbs against proteins and peptides and not for ca ...

Effective multi-agent/multivalent vaccines that confer protection against more than one disease are highly desirable to the patient and to health-care professionals. Electroporation of DNA vaccines, whereby tissues injected with DNA are subjected to localized electrical c ...

Vaccination is historically one of the most important methods for preventing infectious diseases in humans and animals. New insights in the biology of the immune system allow a more rational design of vaccines, and new vaccination strategies are emerging. DNA vaccines have been proposed as a ...

Strategies to improve vaccine efficacy are still required. The immunogenicity of DNA vaccines is strongly improved by electroporation (EP). The skin is populated with a wide variety of immune cells, making it an attractive tissue for vaccine delivery. Here we describe a method for the EP-medi ...

Vaccines to prevent HIV remain desperately needed, but a number of challenges, including retroviral integration, establishment of anatomic reservoir sites, high sequence diversity, and heavy envelope glycosylation. have precluded development of a highly effective vaccine. ...

The development of effective vaccines has helped to eradicate or control the spread of numerous infectious diseases. However, there are many more diseases that have proved more difficult to eliminate using conventional vaccines. The recent innovation of DNA vaccines may provide a “boo ...

Vaccines have evolved for hundreds of years, but all utilize the premise that safely pre-exposing the host to some component of a pathogen allows for enhanced immune recognition, and potential protection from disease, upon encountering the pathogen at a later date. Early vaccination stra ...

Irreversible electroporation has recently been applied to tissue and tumor ablation. This animal model was developed to evaluate optimal parameters for subcutaneous tumor ablation. The immortalized Panc1 tumor cell line was used for generating tumors via subcutaneous injecti ...

Electroporation is a delivery technique that is gaining popularity among the veterinary community due to its low cost, ease of application, and flexibility. It combines the administration of pharmaceutical compounds such as chemotherapy agents, antisense, and plasmids to the appl ...

Delivering genes and other materials directly into the tumor tissue causes specifically localized and powerfully enhanced efficacy of treatments; however, these specific effects can cause rapid, drastic changes in the appearance, texture, and consistency of the tumor. These chan ...

Gene therapy may represent a promising alternative strategy for cardiac muscle regeneration. In vivo electroporation, a physical method of gene transfer, has recently evolved as an efficient method for gene transfer. Here, we describe two protocols involving in vivo electroporati ...

Several of the biological processes involved in the pathogenesis of acute lung injury and acute respiratory distress syndrome after lung contusion are regulated at a genetic and epigenetic level. Thus, strategies to manipulate gene expression in this context are highly desirable not o ...

Gene transfer by electroporation is a versatile technique that can effectively deliver DNA or RNA into almost all types of cells or tissues. As a widely used nonviral approach, electroporation possesses several advantages over viral methods including non-immunogenicity and local t ...

Ocular injection (intravitreal, subretinal, or into the anterior space) is an efficient approach to deliver many classes of drugs, cells, and other treatments to various cell types of the eye. In particular, subretinal injection is efficient since delivered agents accumulate as there is no ...