Phillips Petroleum Company

The Salk Institute Biotechnology Industrial Associates, Inc.

Richard G Buckholz

Michael Miller Harpold

James M Cregg

Juerg F Tschopp

Gregory P Thill

Robert Steven Siegel

Lillian M Wondrack

Thomas S Vedvick

Michael E Engel

Jennifer A Kinney

Mary S Urcan

The present invention discloses the Pichia pastoris acid phospbatase gene, which includes the 5' regulatory region, signal sequence, structural gene, and 3' transcription termination sequence. Also disclosed are methods of using these fragments, which include but are not limited to the secretion of proteins from cells and the regulation of the transcription of DNA. DNA vectors containing the acid phospbatase gene or fragments thereof and hosts transformed with these vectors are also disclosed. Additionally, integrative vectors which direct integration at the Pichia pastoris PHO1 locus and a method of identifying these disruptants is disclosed.

Pichia pastoris acid phosphatase gene, gene regions, signal sequence and

Biologically active aprotinin (APR) molecules, naturally occurring, relatively short, single chain polypeptides, are prepared by growing methylotrophic yeast transformants containing in their genome at least one copy of a DNA sequence operably encoding APR, in operational association with a DNA sequence encoding the S. cerevisiae alpha mating factor pre-pro sequence (including a processing sequence selected from lys-arg-(glu-ala).sub.x, wherein x is an integer falling in the range of 0-3), both under the regulation of a promoter region of a gene of a methylotrophic yeast, under conditions allowing expression of said DNA sequences, and secretion of APR molecules into the culture medium. Also disclosed are novel DNA fragments and novel recombinant yeast strains which are useful in the practice of the present invention.

DNA for expression of aprotinin in methylotrophic yeast cells

Epidermal Growth Factor (EGF) is produced by recombinant DNA technology in Pichia pastoris yeast cells.

Production of epideramal growth factor in pichia pastoris yeast cells

Novel DNA constructs comprising regulatory regions plus the structural coding region for hepatitis B surface antigen (HBsAg) are disclosed. The regulatory regions employed are responsive to methanol, non-catabolite repressing carbon sources and catabolite repressing carbon sources followed by carbon source starvation. The novel constructs are incorporated into a variety of linear and circular plasmids. Such plasmids are used to transform suitable hosts and ultimately used for the production and isolation of hepatitis B surface antigen in high yields.

Yeast production of hepatitis B surface antigen

Growing community of inventors

Encinitas, CA, United States of America

Richard G Buckholz