African Journal of Biotechnology Vol. 5 (24), pp. 2423-2427, 16 December 2006   

ISSN 1684–5315 © 2006 Academic Journals        

Full Length Research Paper

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

Minsheng Wu

Department of Crop Science, North Carolina State University Raleigh, NC 27607, USA. Phone: 919-513-4647. Fax: 919-5157959.

Corresponding author’s E-mail: mwu@ncsu.edu.

Accepted 10 November, 2006

A novel plant differentially expressed gene cloning technique called cDNA-AFLP-SSPAG was described. 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 silver-staining polyacrylamide gels provide a novel and very powerful gene cloning method.

Key words: Cloning, maize, silver-staining polyacrylamide gels, GRF gene, heterosis.