Iain Kilty Phil Vickers
Pfizer Central Research, Sandwich, UK. 01304 618482
Differential display PCR (DDPCR) is a widely used technique for identifying genes differentially expressed between cell types. Application of DDPCR to allergic disease provides a potential means for elucidating the mechanisms involved in allergic responses. In order to successfully apply DDPCR, high-quality, reproducible denaturing polyacrylamide gels are required to fractionate PCR fragments. Stratagenes CastAway system* is convenient and consistently yields the high-quality band resolution that is necessary for DDPCR.
Allergic disease represents a major public health problem in most countries, affecting people of all races. Over 100 million people worldwide suffer from asthma alone.1 Allergic disease is treated predominantly with the use of steroids. Although steroids are effective in eliminating the symptoms of allergic disease, they also cause a number of detrimental side effects. Thus, a greater understanding of the genes involved in the pathogenesis of allergic disease is needed to allow more acceptable drugs to be developed.
As leukocytes play a key role in mediating allergic responses, a comparison
of the expression levels of genes in the leukocytes of people suffering from
allergic disease as compared to nonallergic controls may provide insights into
the basis of such disease. A number of methods may be used for identifying
differentially expressed genes, such as high-throughput sequencing of expressed
sequence tags (ESTs), subtractive hybridization of cDNA and DDPCR. DDPCR is
often the method of choice for identifying differentially expressed genes
because it (1) requires relatively little starting material, (2) can be used to
screen a large proportion of the mRNA population and (3) does not require the
sequencing of large number