Oxidative stress poses a serious problem to the production of biologics via industrial cell lines, with reactive oxygen species (ROS) being the most common oxidant found in cells as it a natural byproduct of aerobic metabolism. The sulfur-containing amino acid methionine has the ability to act as a ROS scavenger, protecting crucial protein residues within cells. This project looked to decrease oxidative stress within S. cerevisiae via the overexpression of the methionine transporter, SLC7A5. The experiment would have served as a proof of concept to be tested in commonly used industrial cell lines, such as CHO. No experimental results were able to be obtained due to unsuccessful gene transformation into S. cerevisiae. This was attributed to issues with the construction of the plasmid containing the SLC7A5 gene. A review of the experimental procedure revealed that a different approach to plasmid construction should be explored if further work were to be done.

• This report represents the work of one or more WPI undergraduate students submitted to the faculty as evidence of completion of a degree requirement. WPI routinely publishes these reports on its website without editorial or peer review.

Creator

• Bhatnagar, Kavim
• LaFortune, Christopher

Publisher

• Worcester Polytechnic Institute

Identifier

• E-project-042822-112429
• 65156

Mot-clé

• Oxidative Stress
• CHO
• Methionine
• Biotechnology
• Genetic Engineering
• Yeast

Advisor

• Young, Eric Mosher

Year

• 2022

Date created

• 2022-04-28

Resource type

• Major Qualifying Project

Major

• Chemical Engineering

Rights statement

• In Copyright