Bioengineered Proteins for Chemical/Biological Defense,Protection, and Decontamination

Period of Performance: 09/03/2002 - 09/02/2004

$500K

Phase 2 STTR

Recipient Firm

Integrated Genomics, Inc.
2201 West Campbell Drive,
Chicago, IL 60612
Principal Investigator
Firm POC

Research Institution

Keck Graduate Institute
535 Watson Drive
Claremont, CA 91711
Institution POC

Abstract

Pichia pastoris is a budding yeast used by hundreds of academic and industrial researchers to synthesize a variety of eukaryotic recombinant proteins. P. pastoris is also a model system used to investigate certain areas of eukaryotic cell biology, such as peroxisome biogenesis and degradation, and Golgi organization. The goals of this project are: (1) to determine, with high fidelity, the DNA sequence of the genome of P. pastoris by a combination of "shot-gun" sequencing and "primer-walking" gap closing; (2) to annotate the expected 6,000 plus ORFs; (3) to develop a publicly accessible web site dedicated to the dissemination of information on the sequencing and annotation results; (4) to construct and make available DNA microarrays focused on ~500 P. pastoris genes whose expression is relevant to recombinant protein production; and (5) to construct certain modified P. pastoris strains for improved production of recombinant proteins. The sequencing, assembly, and preliminary gene annotation work will be performed by Integrated Genomics (IG), an experienced microbial sequencing and analysis company. The hosting of the web site, DNA microarray, and host strain constructions will be performed at the Keck Graduate Institute (KGI). KGI is a new educational and research institution focusing on bioinformatics and genomics technologies. The availability of the DNA sequence of the P. pastoris genome with associated materials and information as proposed in this application will be of enormous benefit to the many commercial and academic researchers utilizing this novel but important yeast. Because of the similarity in amino acid sequence between most P. pastoris proteins and their Saccharomyces cerevisiae counterparts, much of the great wealth of information generated over the years for S. cerevisiae genes and products will be transferred directly to those of P. pastoris. Users of P. pastoris will be able to rapidly identify genes of interest and, in conjunction with PCR methodology, have these genes in hand for further study literally overnight. Thus, the P. pastoris genomic database will be a commercially valuable product in itself. The focused DNA microarrays will be a useful tool to commercial laboratories that wish to monitor gene expression as part of their bioprocess development efforts for new recombinant protein production strains. Finally, the P. pastoris strains constructed for this project are expected to lead to improved levels of recombinant protein production from P. pastoris and therefore, to be valuable to the many companies and academic labs endeavoring to improve the efficiency of their recombinant processes.