Group Final Project

L-Protein Mutants

Problem: How to improve the stability and auto-folding of the lysis protein of an MS2-phage? This mechanism is key to understanding how phages may help address antibiotic resistance.

After going through the readings, including the group final project document a Plan A would be: (This stays within scope, MurJ and multi-target approaches seem intersting though…)

1. Use computational tools like AlphaFold2 or ProteinMPNN to identify mutations that improve intrinsic stability and auto-folding of the lysis protein

2. Target mutations that strengthen the hydrophobic core, eliminate aggregation-prone regions, or introduce stabilising interactions like salt bridges

3. Engineer the lysis protein to fold correctly without requiring DnaJ or any other bacterial chaperone

4. Design mutations that also accelerate oligomerisation or enhance membrane pore-forming activity for faster lysis

5. Synthesise the mutant gene via Twist, clone into plasmid using Gibson Assembly, validate structural integrity with Nuclera, then test in E. coli.