Identification of transcriptional activation domains in ZmMYB40 and ZmMYB95
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Series/Report no.:2014 Richard J. and Martha D. Denman Undergraduate Research Forum. 19th
The phenolic biosynthesis pathway in maize (Zea mays L.) is responsible for the production of phenylpropanoids, complex organic compounds integral to plant secondary cell walls. Lignin, one of the pathway products, compromises efficient conversion of plants to fuel. Our goal is to identify and remove the transcriptional activation domains from MYB40 and MYB95, homologous maize transcription factors believed to act in phenolic pathway regulation. Identifying the domains provides fundamental knowledge, while their removal allows MYB40 and MYB95 to be used as baits in yeast two-hybrid protein-protein interaction analysis. Both MYB40 and MYB95 activate transcription in yeast when fused to the GAL4 DNA-binding domain. This activation, which occurs without a gene of interest in the GAL4 DNA-activating domain, interferes with further analysis by producing false positives. Transcriptional activation domains responsible for this effect were predicted using hydrophobic cluster analysis. Genes were truncated to eliminate the predicted regions and tested for transcriptional activation. Regions of approximately fifty and ninety amino acids were identified to contain transcriptional activation domains in MYB40 and MYB95, respectively. Future work will attempt to identify the exact transcriptional activation domains within these regions, as well as identify and remove areas of similar function in other maize transcription factors. Without these activation domains, protein-protein interaction analysis can be used to elucidate the regulatory network involved in the maize phenolic biosynthesis pathway and establish an information database for use in future studies. This knowledge can be exploited to modify lignin production in fuel crops to make biofuel manufacturing more efficient and cost-effective.
Biological Sciences: 1st Place (The Ohio State University Denman Undergraduate Research Forum)
Academic Major: Chemistry
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