Submitted by Juliana Artier on
Rises on CO2 levels in our atmosphere and continuous populational growth demands new, innovative and clean technologies to be developed for both carbon mitigation and crop enhancement for food. Synthetic biology is a great tool currently being used to modify and bioengineer organisms that are genetically malleable. This allows us to introduce difficult to study pathways from other organisms into model ones.
A large group of bacteria uses the Calvin-Benson-Bassham (CBB) cycle to fix carbon dioxide into sugar. An important part of this process is held inside a microcompartment, the carboxysome, present in cyanobacteria. The most critical enzyme for carbon fixation, Rubisco is housed there, closely to its substrate, CO2, increasing the rate for carboxylation. There’s an interest in understanding different forms of Rubisco, their structure and function, using in vivo strategies, to accommodate for the cellular internal microenvironment, instead of isolated in vitro.
The goal of this project is to use genetic engineering to introduce different Rubisco forms into a model organism Escherichia coli. This will allow us to further study structure, assembly and functionality of these enzymes in vivo. This is an important step that will lay the foundation for the implementation of enhanced carbon fixation into different groups of cyanobacteria and plants.
This project will involve working with a graduate student and a postdoc to clone the rubisco genes into Escherichia coli. Mentees will learn and apply 1) PCR for cloning and genotyping 2) DNA assembly techniques such as Gibson assembly 3) Generation of transgenic Escherichia coli lines using assembled DNA and 4) Analysis of transgenic Escherichia coli using techniques such as growth phenotyping. Additional biochemical techniques, as well as opportunities for independent research projects, may also be incorporated based on mentee interest and as the project develops. It will require basic and build upon student’s genetic engineering and microbiology techniques.
Minimum qualifications: Biology 1A or 1B (may be taken concurrently), 3.0 GPA, and interest in microbiology and genetics. Hours are 10+, and negotiable. Enthusiasm for research is necessary, care-to-detail and commitment to scheduled work times are critical, as well as team work skills.