Improved Tools for Expression of Recombinant Genes in the Yeast Pichia pastoris
DESCRIPTION (provided by applicant): The goal of the proposed research is to identify and develop new basic tools for the expression of recombinant genes for the methylotrophic yeast Pichia pastoris. In recent years this yeast has become an important hostfor the production of biopharmaceuticals with over five thousand publications on its use and at least two human biopharmaceuticals (human serum albumin and ecallantide) on the market in the US and others at various stages in the clinical trial pipeline. This project will be done in part by and wholly for Biogrammatics, Inc. a small biotechnology company incorporated in January of 2008 whose business is to develop, utilize and sell improved tools for heterologous protein expression in this yeast. The business strategy of Biogrammatics involves two general objectives: 1) the sale of novel P. pastoris components and; 2) the construction, testing and improvement of custom recombinant strains of this yeast for customers. Three specific aims are outlined in thisapplication: 1) to conduct a genome wide search for new strong and/or well regulated promoters for expression of recombinant gene in P. pastoris; 2) to develop improved methods for preparation of competent P. pastoris cells for DNA-mediate transformationsand 3) to identify and develop of improved signal sequences to direct the secretion of recombinant proteins from this yeast The project, if funded through a two year Phase I SBIR-SHIFT award mechanism will result in products for Biogrammatics without theneed for a Phase II. At the initiation of this project, the head PI on the application, Dr. Cregg will shift from being a professor at the Keck Graduate Institute to an employee of Biogrammatics. ) PUBLIC HEALTH RELEVANCE: Proteins/peptides are the fastest growing sector in 900 billion dollar therapeutic agent market and hold the promise for true personalized medicine and stem cell therapies. Optimization of Pichia pastoris secretory promoters, transformation efficiency and signal sequences will significantly increase the success rate for recombinant protein production, leading to faster clinical trials and drug approvals in the rProtein therapeutic arena.
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