Aplication of Activation Tagging In Rice For Gene Discovery : Identification and Isolation of Genes Involved in Drought Tolerance
Pelaksana : Satya Nugroho
Following the sequencing of the rice genome, the challenge to the scientific community is to identify functions for each of the predicted 50,000 rice genes.
In an attempt to contribute to that effort, in collaboration with the Plant Research International (PRI), Wageningen, the Netherlands, we are initiating the development of mutant rice library by transposon Ac/Ds insertions containing activation-tag. Activation tagging in plants has been proposed as a novel gene isolation method and has been shown to be effectively applied using T-DNA inserts and the Ac/Ds transposon system. These inserts carry strong activating sequences, from the well-characterized cauliflower mosaic virus (CaMV) 35S enhancer or promoter, which functions as transcriptional activator and can act on genes adjacent to the insertion site and modify their expression by overexpressing the tagged genes to reveal dominant gain-of-function phenotypes. Mutant library are being generated by transforming rice cv Nipponbare with the activation-tag construct. So far, roughly 500 candidate transgenic rice plants carrying the activation tag construct have been generated. This rice lines are being acclimatized in the glass-house and DNA’s are going to be isolated. Insertion analyses are going to be performed to estimate the copy number of the inserted activation tag construct in the genome. The second (T1) generations will be harvested and the activity of the Ds element will be analyzed.
Kata kunci: Gene Discovery, Ac/Ds, activation tag, mutant population., RUTI 2007 P2 Biotek LIPI