Team:UChicago

ChromoVector

A Chromosomal Plasmid for Pichia Pastoris

What is Yeast?

Yeast is well known as the special ingredient in bread. And while that is true, yeast is so much more then that:
Yeast is a fungi, with cells similar to that of humans but can grow a lot faster. In fact they are so closely related, that yeast and human cells share genes. Yeast is used in science to test drugs before testing on humans. Yeast hosts offer the advantages of eukaryotic membrane-targeting machinery, chaperones, and the potential for faithful post translational modifications. Pichia pastoris or methylotrophic yeast, is a type of yeast used for protein production. It has a reproduction rate and can be grow on simple, inexpensive mediums.

What We Created

For the 2018 iGEM competition, the University of Chicago GeneHackers are continuing to develop a centromeric plasmid for Pichia pastors. Centromeric plasmids combine the stability of a chromosome and the flexibility of a plasmid to create the perfect tool for the biotechnical industry. Pichia pastors, a strain of yeast often used in this field, lacks a centromeric plasmid, which limits the research that can be done with this species. Such a plasmid will be useful for a multitude of purposes, including bioengineering and industrial applications. We have found several putative sequences within the genome of Pichia pastoris that will allow our bacterial-yeast shuttle vector to integrate into the yeast's chromosomes. Upon creating our centromeric plasmid, we will perform fluorescence and sectoring assays to ensure the DNA is distributed evenly among daughter cells of the yeast, and we will work to optimize the P. pastors chromosomal sequence. Overall, our goal for this year is to complete our P. pastoris centromeric plasmid.

Project Significance

As stated before, our ned plasmid will be useful for a multitude of purposes, including bioengineering and industrial applications. Todays science focuses on E. Coli because it is cheaper and easier to use. However, yeast is superior then E.Coli because it offers the advantages of eukaryotic membrane-targeting machinery, chaperones, and faithful post translational modifications. With the success of our project, a wide variety of labs and companies can use P. pastors to solve problems.