Undergraduate Thesis Or Project
 

Testing the effectiveness of the λ insulator in hybrid poplar

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  • Genetic engineering techniques allow scientists to create plants with desired traits. However, there is unwanted variability in transgene expression due to enhancer-promoter interactions and the influence from genetic elements near the insertion site of the transgene, which integrates into a random location in the genome. In an attempt to reduce the impact from both enhancers in the construct and in the genome, we investigated the enhancer-blocking function of a 1-kb bacteriophage λ fragment in hybrid poplar (Populus tremula x alba), which is a promising feedstock for biofuel production. The fast growth rate and high cellulose content allow hybrid poplar to produce high levels of biomass within a short period of time, which can then be converted into bioenergy products. Additionally, genetic engineering techniques have enabled scientists to create transgenic poplar trees with various desired traits which could serve to improve performance and lead to higher or more stable yields from bioenergy poplar trees. We placed the λ insulator fragment between a cauliflower mosaic virus 35S promoter (upstream) and a promoter from the meristem-specific gene WUSCHEL (WUS) fused to the β-glucuronidase (GUS) reporter gene on a plant transformation vector. Previous work with the non-insulated construct showed variable GUS expression in non-meristematic tissues, such as leaves. Our results showed that the presence of λ insulator fragment led to a more meristem-specific transgene expression pattern and a decrease in expression levels in both meristems and nonmeristematic tissues.
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  • This project is partially funded by AHB Education project supported by an Agriculture and Food Research Initiative Competitive Grant no. 2011-68005-30407 from the USDA National Institute of Food and Agriculture (NIFA).
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