Invitrogen™ PichiaPink™ Kit zur Sekretionsoptimierung

Beschreibung

The PichiaPink™ Secretion Optimization Kit is a new recombinant protein expression system based on the yeast Pichia pastoris. The PichiaPink™ Secretion Optimization Kit gives you convenient and cost-efficient bioproduction from shake flask to 1000-liter production of your recombinant protein. The PichiaPink™ Secretion Optimization Kit comes with our new Pichia strains, the pPINK-HC and pPINK-LC vectors, and secretion signal sequences for production of your protein.

• Get a complete kit to get started with protein production in Pichia pastoris.
• Screen your Pichia transformants rapidly by auxotrophy and color selection.
• Reduce protein degradation with protease-deficient Pichia pastoris strains.
• Optimize protein secretion with one of 8 secretion signal sequences.

The PichiaPink™ Secretion Optimization Kit comes with:

• PichiaPink™ Vector Kit (Cat. No. A11152)
• PichiaPink™ Secretion Signal Set (A11155)
• PichiaPink™ Expression Strain Set (A11154)
• PichiaPink™ Media Kit (A11156)

A New Way to Screen Pichia Transformants

The PichiaPink™ Secretion Optimization Kit uses adenine auxotrophy to select for Pichia transformants. The PichiaPink™ strains have the ade2 genotype and require complementation with the ADE2 gene to grow without adenine. Use of an auxotrophic marker also makes it easier to maintain transformants. No antibiotics are required to maintain your transformants.

In addition, you can screen for Pichia transformants by color. The untransformed PichiaPink™ strains appear as pink colonies while transformed PichiaPink™ strains appear as white colonies (Fig 1).

New Expression Vectors for Pichia

The PichiaPink™ Secretion Optimization Kit comes with the new PichiaPink™ Vector Kit. The kit includes two vectors, pPINK-LC and pPINK-HC. The pPINK vectors are built around the ADE2 gene for complementing the ade2-deficient Pichia strains. Both vectors use the methanol-induced AOX1 promoter to express your protein. The pPINK-LC integrates at low-copy number into the Pichia genome while the pPINK-HC vector integrates with high-copy number.

Both pPINK-LC and pPINK-HC vectors are compatible with the PichiaPink™ Secretion Signal Set. Using a 3-way ligation, you can add one of 8 secretion sequences to your gene. Note: the pPINK vectors express proteins intracellularly by default.

Both the PichiaPink™ Vector Kit (Cat. No. A11152) and PichiaPink™ Secretion Signal Set (A11155) can be ordered separately to refill the PichiaPink™ Secretion Optimization Kit.

Keep Your Proteins Intact

The PichiaPink™ Secretion Optimization Kit comes with the PichiaPink™ Expression Strain Set. This set contains the 4 Pichia pastoris strains for use with the PichiaPink™ Expression System. All 4 strains have the ade2 genotype. In addition, 3 strains include knockouts in 2 protease genes. Protease-deficient strains reduce the need for protease inhibitors when growing your cells and improve the yield of your protein. Test a single protease knockout or the double protease knockout to find the one that works best for you.

You can order the PichiaPink™ Expression Strain Set (Cat. No. A11154) separately to refill the PichiaPink™ Secretion Optimization Kit.

Media Pouches to Get You Started

The PichiaPink™ Media Kit is a set of convenient prepackaged media pouches for use with the PichiaPink™ Secretion Optimization Kit. The PichiaPink™ Media Kit contains media needed to grow PichiaPink™ strains from frozen stocks to cultures ready for transformation and selection. You can order the PichiaPink™ Media Kit (Cat. No. A11156) separately to refill the PichiaPink™ Secretion Optimization Kit.

Why Choose the PichiaPink™ Yeast Expression System?

The PichiaPink™ Yeast Expression System is based on the yeast Pichia pastoris. Advantages of Pichia pastoris include rapid growth, well-defined genetic background, simple media formulation, and easy handling. For over 30 years, Pichia pastoris has been used by labs around the world for producing hundreds of different proteins from many species including human (Ref 1, 2). The PichiaPink™ Yeast Expression System allows convenient and cost-effective protein production from small to large scales.

For information on obtaining a commercial-use license for the PichiaPink™Yeast Expression System, please inquire at outlicensing@lifetech.com.

References

1. Cereghino JL, Cregg JM. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol Rev. 2000 Jan;24(1):45-66. [PubMed]

2. Cereghino GP, Cereghino JL, Ilgen C, Cregg JM. Production of recombinant proteins in fermenter cultures of the yeast Pichia pastoris. Curr Opin Biotechnol. 2002 Aug;13(4):329-32. [PubMed]