Mutant Lysenin Pores for Lower-Variance Nucleotide Discrimination

The patent applies parameter changes by systematically modifying amino acid residues at specific positions within the pore structure. Mutations are introduced to alter the chemical and physical properties of the pore environment, thereby optimizing the balance between current range expansion and variance reduction. This involves changing parameters such as charge distribution, hydrophobicity, and steric constraints at key positions to achieve improved nucleotide discrimination while maintaining signal stability.

The patent applies local quality by creating distinct functional zones within the pore structure. Specific regions are engineered to interact with particular nucleotide features, allowing the pore to read multiple nucleotides simultaneously while maintaining interpretable signals. This involves assigning different local properties to different segments of the pore, such as varying charge densities or steric constraints at specific positions, to enable multi-nucleotide reading without losing signal interpretability.