James M. Cregg, PhD
Professor EmeritusRecombinant Gene Expression, Recombinant Protein Synthesis, Protein Purification, Peroxisome Biogenesis and Autophagy, Molecular Biology, Biochemistry, Gene Function, Homologous Recombination, Genomics, Bioprocessing, Optimization of Cultures in Fermentors
Dr. Cregg earned his PhD in biology from Rice University in Houston, TX in 1978 and was a postdoctoral fellow at Scripps Clinic and Research Foundation in La Jolla, CA from 1978 to 1980. Previous appointments have been with BRL/Life Technologies in Gaithersburg, MD (1980–1983), the Salk Institute Biotechnology Industrial Associates in San Diego, CA (1983–1989), and the Oregon Graduate Institute in Beaverton, OR (1989–1999). In 1988, Dr. Cregg also spent a year on sabbatical in the Netherlands at the State University in Groningen. Dr. Cregg was one of the founding Professors at KGI and has continued his research on the development and use of yeast, Pichia pastoris, as a system for the production of recombinant proteins, for which he is internationally known. He has also helped found two biotechnology companies, Alder Biopharmaceuticals and BioGrammatics. His work has majorly impacted the development of pharmaceutical proteins such as a chimeric monoclonal antibody, which works against rheumatoid arthritis and certain cancers, research he did in collaboration with Alder. BioGrammatics is a company that specializes in expressing various genes in Pichia pastoris for their contract partners. He has over 100 published papers and holds 14 patents in this area. Dr. Cregg has been invited to give keynote addresses at numerous meetings and serves as a reviewer or executive editor on five peer-reviewed journals in the biotechnology field.
Ternes P, Wobbe T, Schwarz M, Albrecht S, Feussner K, Riezman I, Cregg JM, Heinz E, Reizman H, Feussner I, et al. "Two pathways of sphingolipid biosynthesis are separated in the yeast Pichia pastoris". Biological Chemistry 2011;286(13):11401-11414
Nazarko VY, Nazarko TY, Farre JC, Stasyk OV, Warnecke D, Ulaszewski S, Cregg JM, Sibirny A, Subramani S. "Atg35, a micropexophagy-specific protein that regulates micropexophagic apparatus formation in Pichia pastoris". Autophagy 2011;7(4):375-385
Zhang P, Zhang W, Zhou X, Bai P, Cregg JM, Zhang Y. "Role of two Pichia pastoris hexose transporter homologues in sugar uptake and catabolite derepression (sugar-induction) of AOX1 promoter". Applied and Environmental Microbiology 2010;76:6108-6118
Cregg JM, Tolstorukov I, Kusari A, Sunga J, Madden K, Chappell T. "Expression in the Yeast Pichia Pastoris". Guide to Protein Purification, Second Edition 2009;463:169-189
Wriessnegger T, Sunga AJ, Cregg JM, Daum G. "Identification of phosphatidylserine decarboxylases 1 and 2 from Pichia pastoris". Fems Yeast Research 2009;9(6):911-922
Sunga AJ, Tolstorukov I, Cregg JM. "Posttransformational vector amplification in the yeast Pichia pastoris". Fems Yeast Research 2008;8(6):870-876
Hartner FS, Ruth C, Langenegger D, Johnson SN, Hyka P, Lin-Cereghino GP, Lin-Cereghino J, Kovar K, Cregg JM, Glieder A. "Promoter library designed for fine-tuned gene expression in Pichia pastoris". Nucleic Acids Research 2008;36(12):e76
Wriessnegger T, Gubitz G, Leitner E, Ingolic E, Cregg J, de la Cruz BJ, Daum G. "Lipid composition of peroxisomes from the yeast Pichia pastoris grown on different carbon sources". Biochimica et Biophysica Acta-Molecular and Cell Biology of Lipids 2007;1771(4):455-461
Klionsky DJ, Abeliovich H, Agostinis P, Agrawal DK, Aliev G, Askew DS, Baba M, Baehrecke EH, Bahr BA, Ballabio A, et al. "Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes". Autophagy 2008;4(2):151-175
Cregg JM (Ed.) Methods in Molecular Biology. 2nd. 2007. Totowa, NJ., Humana Press. Pichia Protocols.
Cregg JM. "Introduction: Distinction Between Pichia pastoris and Other Expression Systems". In: Cregg JM, editor. Methods in Molecular Biology. 2nd ed. Totowa, NJ.: Humana Press; 2007;389:1-10
Cregg JM. "DNA-Mediated Transformation". In: Cregg JM, editor. Methods in Molecuar Biology. 2nd ed. Totowa, NJ.: Humana Press; 2007;389:27-42
Tolstorukov I, Cregg, JM. "Classical Genetics". In: Cregg JM, editor. Methods in Molecular Biology. Totowa NJ.: Humana Press; 2007;389:189-201
Leon S, Zhang L, McDonald WH, Yates J, Cregg JM, Subramani S. "Dynamics of the peroxisomal import cycle of PpPex20p: ubiquitin-dependent localization and regulation". Journal of Cell Biology 2006;172(1):67-78
Stasyk OV, Stasyk OG, Mathewson RD, Farre JC, Nazarko VY, Krasovska OS, Subramani S, Cregg JM, Sibirny AA. "Atg28, a novel coiled-coil protein involved in autophagic degradation of peroxisomes in the methylotrophic yeast Pichia pastoris". Autophagy 2006;2(1):30-8
Lin-Cereghino GP, Godfrey L, de la Cruz BJ, Johnson S, Khuongsathiene S, Tolstorukov I, Yan MD, Lin-Cereghino J, Veenhuis M, Subramani S, et al. "Mxr1p, a key regulator of the methanol utilization pathway and peroxisomal genes in Pichia pastoris". Molecular and Cellular Biology 2006;26(3):883-897
Research Synopsis
Dr. Cregg's research is aimed at understanding and improving the P. pastoris expression system. P. pastoris can grow rapidly on inexpensive media and offers a subcellular environment conducive to the folding of eukaryotic proteins with the ability to perform post-translational modifications such as folding, proteolytic processing, disulfide bridge formation and glycosylation. A major advantage of the P. pastoris system is the promoter used to control the function of foreign genes is derived from the methanol-regulated alcohol oxidase I gene (AOX1) and is among the most powerful and tightly regulated promoters known. The basic molecular biology of the promoter is being studied with the goal of developing stronger versions of the AOX1 promoter and P. pastoris strains that are able to hyper-induce expression from the promoter. Dr. Cregg is also examining P. pastoris as a model system to investigate the molecular mechanisms controlling the assembly of subcellular organelles known as peroxisomes. P. pastoris is an ideal model system because its peroxisomes contain several key enzymes required for methanol metabolism and the peroxisomes become unusually large and numerous when Pichia is grown on methanol. Mutants of the yeast with defects in the assembly of the organelles are not able to grow on methanol but can grow on other carbon sources. Dr. Cregg and other investigators have utilized these P. pastoris mutants, called pex mutants, to clone approximately 20 PEX genes which with their protein products have provided critical insights into how peroxisomes function. In addition, the amino acid sequences of the yeast PEX proteins have been utilized to search human DNA sequence databases and identify the human PEX gene equivalents, many of which have turned out to be genes affected in patients with Zellweger syndrome, a family of lethal genetic diseases characterized by defects in the assembly of peroxisomes.
Current Research Projects
At present, Dr. Cregg's laboratory is producing several recombinant proteins, including phytase enzyme to be used as a nutritional additive in animal feeds, and galactosidases which will be used as food supplements to reduce or eliminate lactose intolerance in milk and other products.
Future Research Interests
Dr. Cregg's laboratory is looking for collaborators in academia or industry interested in expressing new genes in Pichia pastoris. His group is capable of gene synthesis, gene insertion into an appropriate vector and into P. pastoris, and gene product identification and evaluation. Dr. Cregg can also aid investigators with product purification and development of large-scale production processes.
Candida tropicalis transformation system, patent 5,204,252. Issued April 20, 1993, James M. Cregg, inventor
Transformation of yeasts of the genus pichia, patent 4,879,231. Issued November 7, 1989, James M. Cregg, inventor
Genes from pichia histidine pathway and uses thereof, patent 4,885,242. Issued December 5, 1984, James M. Cregg, inventor
Autonomous replication sequences for yeast strains of the genus pichia, patent 4,837,148 Issued June 6, 1989, James M. Cregg, inventor
Site selective genomic modification of yeast of the genus pichia, patent 4,882,279 Issued November 21, 1989, James M. Cregg, inventor
Pichia pastoris argininosuccinate lyase gene and uses thereof, patent 4,818,700 Issued April 4, 1989, James M. Cregg, inventor
Yeast production of hepatitis B surface antigen, patent 4,895,800 Issued January 23, 1990 James M. Cregg, inventor
Pichia transformation, patent 4,929,555 Issued May 29, 1990, James M. Cregg, inventor
Pichia pastoris alcohol oxidase II regulatory region, patent 5,032,516 Issued July 16, 1991, James M. Cregg, inventor
Strains of pichia pastoris created by interlocus recombination patent 5,122,465 Issued June 16, 1992, James M. Cregg, inventor
Cultures of yeast of the genus Pichia altered by site selective genomic modification patent 5,135,868 Issued August 4, 1992, James M. Cregg, inventor
DNA encoding the alcohol oxidase 2 gene of yeast of the genus Pichia patent 5,166,329 Issued November 24, 1992, James M. Cregg, inventor
Promoter for the Pichia pastoris formaldehyde dehydrogenase gene FLD1 patent 6,730,499 Issued May 4, 2004, James M. Cregg, inventor
Formaldehyde dehydrogenase genes from methylotrophic yeasts patent 20030215920 Filed November 20, 2003 James M. Cregg, inventor
Contact Information
| James M. Cregg, PhD | ||
| Location: | Building 535, Room 21 | |
| Phone: | (909) 607-8562 | |
| james_cregg[at symbol]kgi.edu | ||