Prokaryotic expression is mature and easy to be operated, but when it comes to the proteins of eukaryotic organism, error folding is occurred frequently since glycosylation and folding partner are involved. Fortunately,the expression system of pichiapastorisis complementary to this situation to a certain extent. Relying on powerful synthetic biology technology platform and industry-leading codon optimization software, LifeSct 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.
|Process||Discription||Delivery Form||Turnaround Time|
|1. Express vector construction||First, according to your specific requirements, we will analyze the gene sequence of protein of interest. Through the industry-leading Codon optimization software, the amount of gene expression can be increased by several to tens of times. Then, gene synthesis was conducted and the recombinant plasmids are verified by sequencing to see whether the sequences are 100% correct. LifeSct 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 LifeSct to carry on the expression test.||1 tube of freeze dried plasmid (4ug/tube) and 1 tube of glycerol containing the plasmid TOP10||10-20 business days|
|2. Linearized vector and electricity transforming into pichiapastoris||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 pichiapastoris. After the screening, we select at least 6 colonies for expression laboratory scale test.||screening report of positive colonies||5 business days|
|3. 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 blotto (optional, the customer provides a varified first antibody) to analyze the expression effect.||A copy of laboratory scale test verification report||5 business days|
|4. Amplified cultivation,expression and purification interest protein||To optimize and obtain the strains with appropriate expression conditions and use 1L or greater culture scale 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 blotto (optional, the customer provides a varified first antibody) to verify the accuracy.||1-10 mg recombinant proteins and purification report data||10-15 business days|
|5. Protein post processing (optional)||For those proteins with further special requirements, we 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.||Inquiry||Inquiry|
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