Poster: Late and Moved Abstracts

Abs # 948: Establishment of tag end sequence database from T-DNA activation tagging mutants

Presenter:	An, Suyoung , swim514@postech.ac.kr
Authors	An, Suyoung  (A)   Park, Sunhee  (A)   Kim, Sung-Ryul  (A)   Park, Ki-Kyung  (A)
Affiliations:	(A): laboratory of plant functional genomics

We have previously reported generation of T-DNA activation tagging mutants in japonica rice. By employing the iPCR method, we have isolated tagged end sequences (TES) of the mutant lines at 63% efficiency. The TES database is open to public at www.postech.ac.kr/life/pfg/risd/. Seeds are available upon request. On March 2004, we have accumulated over 7000 TES from activation tagging lines. Approximately a half of the T-DNA was inserted into genic regions without any bias toward nucleotide sequences. Among the 3500 insertions in the intergenic regions, about 40% of them increased expression of the nearby genes. The enhancing efficiency was higher when the enhancer was located near the promoter region. However, the enhancer could elevate a gene expression from 10 kb away, demonstrating usefulness of the system for gain-of-function mutagenesis. We also established DNA pools from 60,000 tagging lines. DNA pools of 1000 lines are adequate for screening a T-DNA insertion within a 2-kb region, although high GC contents reduce the screening efficiency. Chance of finding a T-DNA insertion in a given gene is currently approximately 50%. The pools can also be used for isolation of insertional mutants by transposons such as Tos17 or MITE. The mutant screening service is available. Please email to genean@postech.ac.kr for detailed instruction of primer and probe preparation.