Human POP2 Expression Systems
Optimization of human POP2 protein expression in E. coli, with plans to expand to yeast expression systems for post-translational modifications and enhanced protein functionality.
The Question
What are the optimal conditions for human POP2 protein expression in E. coli in terms of temperature, induction time, and yield?
The Approach
Plasmid-based transformation with time-course and temperature optimization analysis in E. coli BL21(DE3) cells, with planned expansion to yeast expression.
Key Finding
Optimal POP2 expression achieved at 37°C with 6-hour induction, yielding approximately 735 μg of protein.
Research Overview
Our analysis of E. coli expression systems reveals important insights for optimizing human POP2 protein production, with plans to expand to yeast-based systems.
E. coli Expression System Optimization
We designed a comprehensive methodology to optimize protein expression in E. coli, focusing on temperature conditions and induction timing. Future work will expand to yeast expression systems to facilitate post-translational modifications essential for proper protein function.
Research Background
Alzheimer's disease is a major cause of cognitive decline in the U.S. with few effective treatments. Our work was inspired by groundbreaking research on neuroinflammation using nanoligamers at Colorado State University.
Citation
Sharma, N., et al. (2024). "Novel nanoligamer approach for targeting neuroinflammation in Alzheimer's disease." Journal of Neuroinflammation, 21(1), 182-195.
View original researchExpression System
E. coli BL21(DE3) strain with pET28a vector for high-yield recombinant protein expression.
Experimental Design
Plasmid-based transformation with time-course and temperature optimization analysis at 30°C, 37°C, and 40°C.
Mia preparing bacterial cultures for the transformation experiment
Key Findings
Our research reveals important insights about optimizing human POP2 expression in E. coli under various conditions.
Expression Optimization Results
Expression Kinetics
E. coli reaches peak expression (80%) at 24 hours, with optimal induction at 6 hours.
Temperature Optimization
37°C is optimal for E. coli (90% efficiency), while 30°C shows moderate efficiency (75%) and 40°C shows reduced efficiency (45%).
POP2 Expression
Successful expression of human POP2 at 14 kDa, with highest yield at 37°C after 6 hours of induction (~735 μg).
Research Team
Ten passionate STEM students from diverse disciplines collaborated on this groundbreaking research project.
Our Story
This collaborative project combines expertise from biology, mathematics, and computer science to tackle complex research challenges in protein expression.
Diverse Expertise
Our team includes students from biomedical sciences, computer science, mathematics, and economics, bringing multiple perspectives to the research.
Key Contributions
Jacob Leavitt led the project, with lab work by Mia Papantonio, Elena Tran, and Van Mai, and data processing by Mia Krause and Edward.
Conference Resources
Access our research materials to learn more about our findings.
Colorado State University • Department of Biochemistry and Molecular Biology