Bioinformatics-based assessment of the relevance of candidate genes for mutation discovery

Abstract

The bioinformatics resources provide a wide range of tools that can be applied in different areas of mutation screening. The enormous and constantly increasing amount of genomic data obtained in plant-oriented molecular studies requires the development of efficient techniques for its processing. There is a wide range of bioinformatics tools which can aid in the course of mutation discovery. The following chapter focuses mainly on the application of different tools and resources to facilitate a Targeting-Induced Local Lesions in Genomes (TILLING) analysis. TILLING is a technique of reverse genetics that applies a traditional mutagenesis to create DNA libraries of mutagenised individuals that are then subjected to high-throughput screening for the identification of mutations. The bioinformatics tools have shown to be useful in supporting the process of candidate gene selection for mutation screening. The availability of bioinformatics software and experimental data repositories provides a powerful tool which enables a process of multi-database mining. The existing raw experimental data (genomicsrelated information, expression data, annotated ontologies) can be interpreted in terms of a new biological context. This may help in selecting the proper candidate gene for mutation discovery that is controlling the target phenotype. The mutation screening using a TILLING strategy requires a former knowledge of the full genomic sequence of the gene which is of interest. Depending on whether a fully sequenced genome of a particular species is available, different bioinformatics tools can facilitate this process. Specific tools can be also useful for the identification of possible gene paralogs which may mask the effect of mutated gene. Bioinformatics resources can also support the selection of gene fragments most prone to acquire a deleterious nucleotide change. Finally, there are available tools enabling a proper design of oligonucleotide primers for the amplification of a gene fragment for the purpose of mutation screening.