Tilling and Fox-Hunting: New Methods for Functional Analysis of Genes

Theoretical and experimental bases of TILLING and FOX-hunting, new tools for precise identification of genes in functional studies are reviewed. TILLING (Targeting Induced Local Lesions IN Genomes) is a technique combining chemical mutagenesis with its sensitive and precise detection. The method involves PCR amplification of DNA samples pooled following extraction from a collection of chemically-treated organisms and a subsequent screening for mutations using Cel1 endonuclease, detecting mismatches in heteroduplexes [52]. FOX-hunting (Full-length cDNA Over-eXpressing gene hunting system) is a new method of plant gene overexpression, which enables a quick gene isolation and sequencing, paralelly with functional studies.