Session Number
Project ID: CHEM 01
Advisor(s)
Dr. Angela Ahrendt, Illinois Mathematics and Science
Dr. Sarah Stainbrook, Washington University in St Louis
Discipline
Arts & Humanities
Start Date
20-4-2022 8:50 AM
End Date
20-4-2022 9:05 AM
Abstract
Rising global temperatures due to climate change threaten the productivity of key agricultural staple crops such as maize corn, thus having negative implications for food security and production. Increased heat stress impairs plant function by decreasing the efficiency of rubisco activase (RCA), a critical enzyme responsible for net photosynthesis. To address this issue, we performed Ancestral Sequence Reconstruction (ASR) on the RCA gene sequence from Zea maize to form a mutated sequence with improved thermostability. ASR is a technique which uses software to generate a most likely ancestor to a protein sequence, which is often more stable than the wild type. Through ASR, it is possible to enhance the thermostability, promiscuity, and activity of proteins. Several rounds of ASR were performed with different combinations of Archaeplastida species, including those genetically furthest and closest from maize on the phylogenetic tree. From this, two generated sequences were synthetically constructed by a commercial lab. The sequences were cloned into an expression vector, and the protein variants will be expressed, purified, and analyzed for improved heat tolerance compared to the maize wild-type RCA.
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Ancestral Sequence Reconstruction of Rubisco Activase from Zea Maize to Improve Thermal Stability
Rising global temperatures due to climate change threaten the productivity of key agricultural staple crops such as maize corn, thus having negative implications for food security and production. Increased heat stress impairs plant function by decreasing the efficiency of rubisco activase (RCA), a critical enzyme responsible for net photosynthesis. To address this issue, we performed Ancestral Sequence Reconstruction (ASR) on the RCA gene sequence from Zea maize to form a mutated sequence with improved thermostability. ASR is a technique which uses software to generate a most likely ancestor to a protein sequence, which is often more stable than the wild type. Through ASR, it is possible to enhance the thermostability, promiscuity, and activity of proteins. Several rounds of ASR were performed with different combinations of Archaeplastida species, including those genetically furthest and closest from maize on the phylogenetic tree. From this, two generated sequences were synthetically constructed by a commercial lab. The sequences were cloned into an expression vector, and the protein variants will be expressed, purified, and analyzed for improved heat tolerance compared to the maize wild-type RCA.