Prokaryotic expression is mature and easy to be operated, but when it comes to the proteins of eukaryotic organism, error folding is a frequent occurrence since glycosylation and folding partner are involved. Fortunately,the expression system of pichia pastorisis is complementary to this situation to a certain extent. Relying on powerful synthetic biology technology platform and industry-leading codon optimization software NGTM Codon, Synbio Technologies provides you high-quality and considerate yeast expression protein services with more than 10 years’ experience in protein expression and purification, which can effectively save your cost and time.
Expression vector construction (optional)
First, according to your specific requirements, we will analyze the gene sequence of protein of interest. Through the industry-leading NGTM Codon optimization software, the amount of gene expression can be increased by several times. Then, gene synthesis is conducted and the recombinant plasmids are verified by sequencing to see whether the sequences are 100% correct. Synbio Technologies provides the service of directly cloning genes into commercial expression vectors such as methanol-induced pPICZα, pPIC9K, or component pGAPZα, etc.. In addition, you can also choose the peculiar yeast special expression vector of Synbio Technologies to carry on the expression test.
Delivery contents: 1 tube of freeze dried plasmid (2~5 µg/tube) and 1 tube of glycerol containing the plasmid TOP10
Delivery TAT: 10-20 business days
Linearized vector andelectricity transforming into Pichia pastoris
You can also give us a constructed recombinant plasmid (please confirm the codon of the sequence is optimized). For those verified recombinant plasmids, we choose suitable enzymes to carry out the linearization and adopt the electric transfer methodto transform them into the Pichia pastoris. After the screening, we select at least 6 colonies for expression laboratory scale test.
Delivery contents: screening report of positive colonies
Delivery TAT: 5 business day
Optimization of expression conditions
During the process of laboratory scale test, we will optimize and test the culture conditions which are suitable for various strains, including temperature, culture medium, inducer type, induction time, induction quantity, and other factors. It is tacitly approved that we use SDS-PAGE or Western Blot (optional, the customer provides a varified first antibody) to analyze the expression effect.
Delivery contents: A copy of laboratory scale test verification report
Delivery TAT: 5 business days
Amplified cultivation,expression and purification interest protein
To optimize and obtain the strains with appropriate expression conditions, 1L or greater culture scale is used to induce recombinant yeast expression. After the induction, one step or multi-step purification is conducted. We use SDS-PAGE to verify the purification and Western Blot (optional, the customer provides a varified first antibody) to verify the accuracy.
Delivery contents: 1-10 mg recombinant proteins and purification report data
Delivery TAT: 10-15 working days
For proteins with tags, you can choose remove service (optional).
Protein post processing (optional)
For those proteins with further special requirements, Synbio Technologies provides you a variety of post processing options including protein tag removal after purification, filtration sterilization, endotoxin removal, freeze-dry, N-terminal sequencing, and other services. Our experienced engineers can provide tailor-made post servicesfor your special needs.
Delivery contents: inquiry
Delivery TAT: inquiry