Development of a simple and powerful method, cDNA AFLP-SSPAG, for cloning of differentially expressed genes

Abstract

A novel plant differentially expressed gene cloning technique called cDNA-AFLP-SSPAG was descrybed. Using this method, Differential mRNA expression was found between high and low heterosis groups of maize. Differential cDNAs were easily obtained from silver stained cDNA-AFLP separated on polyacylamide gels. The cDNA was then reamplified, cloned and fragments were sequenced. Sequenced clones were used as probes in northern dot blot analyses and library screening. Full-length cDNA was cloned from a library suggesting that the cDNA-AFLP technique combined with silverstaining polyacrylamide gels provide a novel and very powerful gene cloning method.