A gene machine for rice
2003
Upadhyaya, N.M. | Zhou, X.-R. | Zhu, Q.-H. | Eamens, A. | Ramm, K. | Wu, L. | Sivakumar, R. | Kumar, S. | Narayanan, K.K. | Thomas, G. | Kato, T. | Yun, D.-W. | Peacock, W.J. | Dennis, E.S.
Insertional mutants will have a crucial role in identifying the function of each of the expected 25,000-40,000 plant genes. We are using the two-component Ac/Ds transposon gene or enhancer trap system, initially delivered through T-DNA, to produce libraries of insertional mutants in rice. Large numbers of Ac or Ds (enhancer or gene trap) transgenic lines have been produced. Mutagenic populations containing Ac and Ds were produced by crossing, cotransformation, or super transformation. In the first screening population, 4.6% of the plants were putative stable Ds trap lines (unlinked to Ac), with 40% containing Ds elements unlinked to Ds donor sites. The regions flanking the original Ds launching pad (T-DNA tag) and Ds transpositions are being sequenced. Among 12 T-DNA tags rescued, 5 were inserted into known sequences. Among 84 Ds tags, 13 were transposed into known sequences, 52 into unknown sequences, and the other 19 in and around the original Ds launching pad. New constructs with bar as a strong selectable marker for Ds itself and for Ds excision as well as a negative selector (tms2) for Ac have been developed for high-throughput screening. Our aim is to build a substantial "gene machine." A labnote-cum-relational database, a tagged sequence database, and a Web site have been set up for efficient handling, processing, and exchange and/or release of tagged sequence data.
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