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From 2010.igem.org

• Home
• Project
  • About The Toolkit
  • Oursystem
  • Producing The Antigen
  • Future Applications
  • Safety
• Toolkit
• Why Pichia
  • Pichia Vs Saccharomyces
  • Easy and Inexpensive to Culture
  • High production of foreign Protein while low levels of endogenous protein
  • Pichia pastoris versus Esherichia coli
• Protocols
  • Preparation Of Electrocompetent Cells
  • Preparation of Heat Shock Competent Cells
  • Transformation Using Electroporation
  • Transformation Using Heat Shock
• The Team
• Notebook

Pichia Tool Kit

New for iGEM 2010 is the groupparts tag.

Welome to the Pichia Tool Box!!! Here you will find a selection of parts designed to facilitate your specific application of Pichia Pastoris!

Pichia pastoris has quickly gained attention as a powerful expression system in the last two decades. Researchers have successfully produced high yields of hundreds of heterologous protein from all orders of life. However, this incredible resource is yet to be tapped into by iGEM! We believe iGEM can play an enormous role in both the application and improvement of this expression system. In our tool box, we have standardized some of the most powerful, reliable and effective parts used today in P. pastoris. From these parts, the ground work is laid for teams to get a head start to optimizing their particular application. We encourage you to experiment, explore and improve this system!

Part Design: Using primers that contain Biobrick ends, parts were isolated from Pichia pastoris chromosomal DNA with PCR reactions. To increase the likelihood of gaining a successful product, a systematic approach was applied for primer design. Two sets of primers were constructed for each part using IDT®’s PrimerQuest and Oligoanalyzer programs. The first set added an Xpa I site on the forward and a Spe I site on the reverse. The second set added EcoRI, Not I and Xba I sites on the forward and Spe I, Not I and Pst I sites on the reverse. In this way, hybrid primer combinations were assembled providing a total of four PCR products. With this system, we have successfully acquired a biobrick prospective product for each part.

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Note that if you want to document a part you need to document it on the [http://partsregistry.org Registry], not on your team wiki.

<groupparts>iGEM010 Georgia_State</groupparts>