Presentation Title

Tracking Gene Regulation Triggered by Environmental Stress within Nuclei of Rice

Start Date

November 2016

End Date

November 2016

Location

HUB 302-77

Type of Presentation

Poster

Abstract

Development and physiological processes within plants are affected by environmental stresses, such as drought and flooding. Rice (Oryza sativa) is a common staple food throughout the world. Further crop production is necessary to sustain the world’s growing population and extreme weather conditions are huge obstacles against productivity. To further explore the plasticity of rice during droughts and floods, advanced methods are being used to make it easier to study cells from the roots and shoots that have specific functions. INTACT (Isolation of Nuclei TAgged in Specific Cell Types) allows isolation of nuclei, using a nuclear-targeting protein which allows access to chromatin (chromosomes containing genes) and nuclear mRNA. This technology can be targeted to specific types of cells. In this project, INTACT transgenic lines were generated and confirmed by the observation of Green Fluorescent Protein (GFP) expression in specific cell types. Validation of the site of insertion of the INTACT construct was conducted using Polymerase Chain Reaction (PCR). INTACT followed by Western Blotting was used to compare three versions of the INTACT nuclear targeting protein. It was shown that the version that anchors the tag through the nuclear envelope is more efficient than the original version that associates with the nuclear envelope. Current experiments aim to estimate the loss of the nucleoplasm, if any, in the INTACT procedure by evaluation of retention of specific protein. These methods enable detailed evaluation of gene regulatory dynamics in multicellular organisms.

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Nov 12th, 1:00 PM Nov 12th, 2:00 PM

Tracking Gene Regulation Triggered by Environmental Stress within Nuclei of Rice

HUB 302-77

Development and physiological processes within plants are affected by environmental stresses, such as drought and flooding. Rice (Oryza sativa) is a common staple food throughout the world. Further crop production is necessary to sustain the world’s growing population and extreme weather conditions are huge obstacles against productivity. To further explore the plasticity of rice during droughts and floods, advanced methods are being used to make it easier to study cells from the roots and shoots that have specific functions. INTACT (Isolation of Nuclei TAgged in Specific Cell Types) allows isolation of nuclei, using a nuclear-targeting protein which allows access to chromatin (chromosomes containing genes) and nuclear mRNA. This technology can be targeted to specific types of cells. In this project, INTACT transgenic lines were generated and confirmed by the observation of Green Fluorescent Protein (GFP) expression in specific cell types. Validation of the site of insertion of the INTACT construct was conducted using Polymerase Chain Reaction (PCR). INTACT followed by Western Blotting was used to compare three versions of the INTACT nuclear targeting protein. It was shown that the version that anchors the tag through the nuclear envelope is more efficient than the original version that associates with the nuclear envelope. Current experiments aim to estimate the loss of the nucleoplasm, if any, in the INTACT procedure by evaluation of retention of specific protein. These methods enable detailed evaluation of gene regulatory dynamics in multicellular organisms.