PichiaPink™ Secreted Protein Kit

Catalog number: A11151

Invitrogen™  Related applications: Protein Expression

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Description

The PichiaPink™ Secreted Protein Expression Kit is a new recombinant protein expression system based on the yeast Pichia pastoris. The PichiaPink™ Secreted Protein Expression Kit gives you convenient and cost-efficient bioproduction from shake flask to 1000-liter production of your recombinant protein. The PichiaPink™ Secreted Protein Expression Kit comes with our new Pichia strains, the pPINK-HC and pPINK-LC vectors, and secretion signal sequences for production of your protein.
  Get started with secreted protein production in Pichia pastoris.
•   Screen your Pichia transformants rapidly by auxotrophy and color selection.
•   Reduce protein degradation with protease-deficient Pichia pastoris strains.

The PichiaPink™ Secreted Protein Expression Kit comes with:
•   PichiaPink™ Secreted Protein Vector Kit (Cat. No. A11153)
•   PichiaPink™ Expression Strain Set (Cat. No. A11154)
•   PichiaPink™ Media Kit (Cat. No. A11156)

A New Way to Screen Pichia Transformants
The PichiaPink™ Secreted Protein Expression 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).

Secrete Your Proteins with the pPINKα-HC Vector

The PichiaPink™ Secreted Protein Expression Kit comes with the pPINKα-HC vector. The pPINKα-HC vector has the α-mating factor signal sequence built-in and allows your protein to be secreted into the media. Secreting your protein into the media makes harvesting and purifying your protein easier. The pPINKα-HC vector is built around the ADE2 gene for complementing the ade2-deficient Pichia strains. This vector uses the methanol-induced AOX1 promoter to express your protein. Further, the pPINKα-HC vector is designed to be integrated at high-copy-number in your Pichia transformants.

The PichiaPink™ Secreted Protein Vector Kit (Cat. No. A11153) can be ordered separately to refill the PichiaPink™ Secreted Protein Expression Kit.

Keep Your Proteins Intact
The PichiaPink™ Secreted Protein Expression Kit comes with the PichiaPink™ Expression Strain Set. This set contains the 4 Pichia pastoris strains for use with the PichiaPink™ Yeast 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™ Secreted Protein Expression Kit .

Media Pouches to Get You Started

The PichiaPink™ Media Kit is a set of convenient prepackaged media pouches for use with the PichiaPink™ Secreted Protein Expression 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™ Secreted Protein Expression 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.

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.

Related Links
Learn more about the PichiaPink™ Yeast Expression System.
Learn more about our other protein expression systems.

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]
For Research Use Only. Not for use in diagnostic procedures.

Figures

Specifications

Bacterial or Yeast Strain: PichiaPink™
Expression: Yeast
Expression Competent Cell: Pichia pastoris strains
Expression Mechanism: Cell-Based Expression
Expression Stability: Stable
Key Function: Secreted Expression
Product Size: 1 kit
Species: P. pastoris
Shipping Condition: Room Temperature

Contents & storage

The PichiaPink™ Secreted Protein Expression kit consists of:
• The PichiaPink™ Secreted Protein Vector kit containing the high-copy number plasmid with the S. cerevisiae Alpha mating factor presequence to yield secreted protein expression.
• The PichiaPink™ Expression Strain set containing 4 glycerol strains
• The PichiaPink™ Media Kit containing 4 basic media.

Documents

Manuals & protocols

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