Differential display of protein tyrosine kinase cDNAs from human fetal and adult brains
Okada, M.; Yamaga, S.; Yasuda, S.; Weissman, S.M.; Yasukochi, Y.
Biotechniques 32(4): 856 858 860 863-5
2002
ISSN/ISBN: 0736-6205 PMID: 11962607 Document Number: 547309
Here we describe a differential display method for surveying the expression of most protein tyrosine kinases and applying it to cDNAs from human fetal and adult brains. The method involves two selective steps for processing the mRNA. At each step, degenerate oligonucleotide primers derived from highly conserved regions of the catalytic domain of the kinases are used. In the display with BstYI and BsiHKAI digests of the cDNA, 65% and 59% of a total of 72 and 63 bands, respectively, represented fragments from a total of 27 different tyrosine kinases. The expression levels of the kinases in the display were comparable with those measured by RT-PCR. This method offers a relatively specific way to display differentially expressed gene families in any tissue and cell type.