[20] Analysis of altered gene expression by differential display

Publisher Summary This chapter presents the analysis of altered gene expression by differential display. Temporal and spatial expression of the 100,000 different genes in a mammalian cell I must be highly regulated and tightly controlled as specificity determines life processes from the fertilized egg to death of the organism. The course of normal development as well as the pathological changes that arise in diseases such as cancer, whether caused by a single gene mutation or complex multigene effects, are driven by changes in gene expression. A fingerprinting technique for mRNAs would be very useful, for instance, by 2D electrophoresis, similar to 2D protein separation. A novel method named "differential display" (DDRT-PCR), according to polymerase chain reaction (PCR) terminology, was developed. This method provides a sensitive and flexible approach to the identification of genes that are differentially expressed at the mRNA level. The method is directed toward the identification of differentially expressed genes, detecting individual mRNA species that are changed in different sets of eukaryotic cells, and then permitting recovery and cloning of their DNA.