Membrane Protein Expression in Pichia Pastoris

The key to the heterologous expression of eukaryotic membrane proteins (EMPs) is to select a suitable expression platform. Pichia pastoris has been proved to be a very versatile one, showing promising results in more and more expression cases. Its special methyl nutrition characteristics combined with the very simple processing of eukaryotic microorganisms with most mammalian-like machines make Pichia pastoris a highly competitive expression system for various proteins with complex functions, and its quantity is compatible with the study of function and structure.

Figure 1. Examples of integral membrane protein structures obtained using protein expressed in P. pastoris. (Pichia pastoris as an expression host for membrane protein structural biology, 2015)

Membrane protein expression based on Pichia Pastoris services is now available at Creative BioMart. Currently, EMPs including (a) Orai Calcium release-activated channel; (b) GIRK2; (c) K2P4.1; (d) AQP2; (e) Histamine H1–T4 lysozyme fusion protein; (f) Monoamine oxidase; (g) Leukotriene synthase; (h) P-glycoprotein has been obtained through our services.

Services Details

Generation and amplification of constructs	Timeline	Deliverable
Protein gene sequence analysis. Codon optimization. Expression vector selection (pPICZ α, pPIC9K and pGAPZα. You can also provide or authorize us to purchase other expression vectors.	2-3 weeks	A. One tube of freeze-dried plasmid B. One tube of glycerol bacteria containing the plasmid TOP10
Linearization vector and electroporation into Pichia pastoris
Recombinant plasmid is linearized after sequence verification. The positive clones are screened by electroporation. At least 6 clones are selected for the expression test.	1 week	Screening report of positive clones
Optimization of expression conditions
Expression conditions test: include temperature, medium, inducer type, induction time and induction amount. SDS-PAGE or Western blot is used to analyze the expression effect by default.	1 week	A small test verification report
Amplification, expression and purification of target protein
The recombinant yeast is induced by 1 L or more culture scale and purified in one or more steps. Purity detection by SDS-PAGE Correctness detection by Western blot.	1-2 weeks	1-10mg recombinant protein and purification report data
Protein post-processing (optional)
We provide a variety of post-processing options, including tag removal, filtration, endotoxin removal, freeze-drying, N-terminal sequencing and other services.

Related Service

Did not find what you need? In addition to the membrane protein expression services, we also offer

• Classic Pichia Pastoris Protein Expression Service for both intracellular and secretory proteins
• Glycoprotein Expression in Pichia Pastoris

Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP.