Protein mutagenesis is a powerful technique used in molecular biology and biochemistry to generate protein variations. This can be used to study protein function, improve protein properties, or even create new proteins with desirable properties. CD Formulation offers fast turnaround protein mutagenesis services to obtain 100% accurate mutant DNA clones. Our team of experts has mature expertise in the field of protein mutagenesis and can customize the design of mutation schemes according to the specific needs of our customers.

What is Protein Mutagenesis?

Protein mutagenesis, a very important DNA manipulation technique, refers to the process of introducing changes or mutations into the gene sequence encoding a protein, resulting in changes in the structure and function of the protein itself. Most protein mutagenesis is based on polymerase chain reaction (PCR), where strand separation is achieved by using a denaturation step to separate complementary strands, and PCR primers can be efficiently polymerized.

Fig. 1 Outline of the novel PCR-based site-directed mutagenesis method. (Rabhi I, et al., 2004)

Explore Our Therapeutic Protein Mutagenesis Service

At CD Formulation, our team provides efficient PCR protein mutagenesis services using the most advanced DNA synthesis technology to ensure the highest accuracy and unlimited mutation sites. Our optimized technology can effectively improve the mutation efficiency and ensure the sensitivity and specificity of the PCR reaction, thereby greatly shortening the experimental cycle and increasing the success rate of the experiment.

In addition, we provide comprehensive upstream and downstream services, including template DNA sequencing, gene synthesis, expression vector construction, and protein expression and purification. Importantly, we can also create your own protein truncation according to your needs.

Site-Directed Mutagenesis Service

Our site-directed mutagenesis services include point or substitution mutations, insertion mutations, and deletion mutations. Our scientists use a targeted PCR-based process to precisely introduce genetic variation and then clone the construct into the vector of your choice. Not only does our site-directed mutagenesis service allow for arbitrary point mutations, insertions, and deletions on plasmids up to 18 kb, but multiple mutation sites can be introduced in a single mutagenesis reaction.

Random Mutagenesis Service

Our random mutagenesis service is based on the introduction of genetic mutations using error-prone PCR and cloning random DNA into the expression vector of your choice. Our random mutagenesis team confirms the desired mutations by DNA sequencing and performs any mutation frequency based on the stability of the construct.

Site-Directed Random Mutagenesis Service

Our site-directed random mutagenesis service involves cloning site-directed randomized DNA into the expression vector of your choice. Similar to our random mutagenesis service, our site-directed random mutagenesis is based on DNA sequencing to confirm the desired mutation and allows for any mutation frequency based on the stability of the construct.

Site Saturation Mutagenesis Service

Our site-saturation mutagenesis service involves cloning DNA into an expression vector or lentiviral vector of your choice. In site-saturation mutagenesis, mutations are introduced at specific locations within a DNA sequence in a controlled and systematic manner. Our team of experts uses different methods, including synthetic oligonucleotides, overlap extension PCR, and oligonucleotide-based PCR to cause these mutations and perform site-saturation mutagenesis. We can perform any targeted mutation frequency, depending on the stability of the construct.

General Workflow for Therapeutic Protein Mutagenesis Service

Fig. 2 Workflow for therapeutic protein mutagenesis. (CD Formulation)

• Designing Mutagenic Primers: According to the target gene sequence, specific mutations such as point mutations, insertions, or deletions are introduced by designing primers. These mutations are usually achieved by specific nucleotide substitutions.
• PCR Amplification: PCR is used to amplify DNA fragments containing mutations. In this step, the template DNA is efficiently replicated, and the use of mutation primers can ensure that the mutation is incorporated into the amplified product.
• Strand Separation and Denaturation: During the PCR process, preheating causes double-stranded DNA to denature and form single-stranded DNA. Then, the primers are bound to the target DNA sequence by cooling.
• Amplification and Mutation: DNA polymerase synthesizes new DNA chains through primer extension to form DNA fragments containing mutations. Multiple rounds of PCR can further increase the expression rate of mutations.
• Cloning and Expression: The mutated DNA fragments are cloned into appropriate expression vectors and then transformed into host cells for protein expression and purification.
• Verification of Mutations: Sequencing and other means are used to verify whether the mutation has been successfully introduced and to ensure that there are no unexpected mutations in the surrounding sequences.
• Functional Analysis: Functional analysis of the expressed protein is performed to evaluate the effect of the mutation on the protein structure and function. This step often involves biochemical experiments, structural biology analysis, etc.

Why Choose Our Therapeutic Protein Mutagenesis Service?

• We have a strong track record in successfully developing therapeutic proteins using protein mutagenesis, ensuring high-quality results.
• We offer comprehensive upstream and downstream services, including template DNA sequencing, de novo gene synthesis, rapid cloning into free expression vectors, custom vector construction, and protein expression and purification.
• Our technology is optimized to introduce point, insertion, and deletion mutations in constructs up to 18 kb, including the target gene and its vector.
• We can perform high-throughput mutagenesis and screening to quickly identify beneficial mutations to accelerate the process of developing and optimizing therapeutic proteins.
• Based on advanced gene synthesis and mutagenesis technologies, we can introduce mutations at any site.
• We provide tailored solutions to meet specific goals, whether it is to improve stability, enhance activity, or modify pharmacokinetics.

Publication

CD Formulation utilizes the latest technologies and protocols to ensure high efficiency and accuracy in its mutagenesis services. Please don't hesitate to contact us if you are interested in our services. We look forward to cooperating with you.