Development of an Efficient Inverse PCR Method for Isolating Gene Tags from T-DNA Insertional Mutants in Rice
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The central goal of current genomics research in plants, as in other organisms, is to elucidate the functions of every gene. Insertional mutagenesis using known DNA sequences such as T-DNA is a powerful tool in functional genomics. Development of efficient methods for isolating the genomic sequences flanking insertion elements accelerates the systematic cataloging of insertional mutants, and thus allows functions to be assigned to uncharacterized genes via reverse genetic approaches. In our current study, we report a rapid and efficient inverse PCR (iPCR) method for the isolation of gene tags in T-DNA mutant lines of rice (Oryza sativa), a model monocot plant.
Key wordsFunctional genomics Gene tag Inverse PCR Rice
The methods described in this report were developed with the support of the Crop Functional Genomic Center, the 21st Century Frontier Program (Grant CG1111); from the Biogreen 21 Program, Rural Development Administration (20070401-034-001-007-03-00); from the Korea Research Foundation Grant funded by the Korean Government (MOEHRD, Basic Research Promotion Fund, KRF-2007-341-C00028); and from Kyung Hee University.