Activation-Tagged and Deletion Mutants and cDNA Microarrays for Functional Genomics of Rice

Kansas State University, Manhattan KS

Investigators

Abstract

Rice genome sequencing projects are defining thousands of new rice genes. However, the critical tools required to determine the functions of these genes are not yet available to public-sector researchers. To remedy this deficiency, novel approaches to generate collections of insertion mutants and to detect mutated genes in a complementary set of rice deletion mutants will be developed. The first objective is to develop and test vectors for creating activation-tagged rice mutants using the Ac/Ds system. Activation tagging allows the identification of gene knockout mutants and mutants exhibiting gene over-expression. The vectors will contain chemical-induced promoters that will allow the controlled transposition of the Ds element. The second objective is to develop and optimize a high throughput protocol for the detection of mutated genes in an existing deletion mutant collection. The strategy is to use polymerase chain reaction (PCR) to detect the mutated genes rapidly in pools of DNA from many mutants. The rapid detection of mutated genes by this protocol will allow the identification of mutants with deletions in specific genes of interest. The international dimension of the project will enrich the training experience of the postdoctoral fellows and extend the impact of the research.