Direct protein evolution is one of the most powerful strategies to create proteins with new or improved properties. Mutagenex provides mutant libraries with a high diversity and accuracy generated by an error-prone PCR and site-directed random mutagenesis (site saturation) at either single or multiple sites.
Site-directed random mutagenesis is also called targeted random mutagenesis or site-saturation mutagenesis. Since most conventional directed evolution strategies, such as error-prone PCR and DNA shuffling, generate large numbers of non-functional mutants rather than functional genes, these approaches inevitably require extensive high-throughput screening systems, which require a high cost and long time.
By targeting specific residues, most codons remain wild-type, which can make mutants maintain the protein backbone's natural structure despite the functional regions' hyper diversity. Therefore, the mutant library can contain a high portion of functional mutants that are more likely to yield positive results.
How can we assist you?
Mutagenex can introduce random or saturated mutations into target sites of genes according to the customer's request with high efficiency of mutagenesis, high accuracy without undesired mutations, and a big library size enough for customer's research.
Applications
Site-directed random mutagenesis has been successful for many directed evolution experiments. The applications of this method include,
| Enzyme | Antibody |
|---|---|
| Alteration of enontioselectivity Alteration substrate specificity New catalytic activity Stability at extreme temperature and pH | Enhanced binding activity and stability Decreased immunogenesity Increased serum half-life |
| Hormone | Other applications |
|---|---|
| Improved association rate or affinity Reduced affinity to increase recycling rate Increased serum half-life | Improvement of cytokines Alteration of GFP fluorescence property DNA or RNA sequence optimization |
The price will be determined depending on target gene size, number of targeting sites, number of target codons (bases or residues) for randomization, total diversity of random mutation, size of primary library and final quality of library.
We provide a mutant library with a high diversity by error-prone PCR as well as site-specific random mutagenesis (site saturation).
To start your site-directed random mutagenesis project, please contact us.