Candida tropicalis transformation system
AssigneeHenkel Research Corporation
InventorsJames M. Cregg, Martin A. Gleeson, Lisa Haas, Stephen Picataggio
Life of this patent
Estimating the term…
Abstract
A transformation system is provided for C. tropicalis, comprising constructs and microorganisms, as well as methods for preparing constructs and microorganisms, and transforming microorganisms. Particularly, a yeast transformation system comprising auxotrophic hosts which are auxotrophic in either an amino acid, purine or pyrimidine pathways and employ DNA constructs comprising genes encoding biosynthetic enzymes which functionally complement the auxotrophies to prototrophies.
Claims9
A method for transforming Candida tropicalis comprising:
- (1) providing an auxotrophic host which requires uracil for growth due to a lack of orotidine-5'-phosphate decarboxylase as the result of mutations in the URA3A and RUA3B genes;
- (2) enzymatically treating the cell wall of said auxotrophic host;
- (3) adding to the resulting spheroplasts a DNA construct comprising the C. tropicalis URA3A or URA3B genes and which functionally compliments the auxotrophic mutation in said host;
- (4) fusing said DNA construct with said auxotrophic host to form transformed cells;
- (5) selecting said transformed cells by their ability to grow in a selective medium lacking uracil.
A transformed Candida tropicalis host derived from C. tropicalis SU-2 which is prototrophic as the result of transformation with either the URA3A or URA3B gene.
A method for transforming Candida tropicalis comprising:
- (1) providing an auxotrophic host which required histidine for growth due to a lack of histidinaol dehydrogenase as the result of mutations in both HIS4 genes;
- (2) enzymatically treating the cell wall of said auxotrophic host;
- (3) adding to the resulting spheroplasts a DNA construct comprising the C. tropicalis HIS4 gene which functionally compliments the auxotrophic mutation in said host;
- (4) fusing said DNA construct with said auxotrophic host to form transformed cells;
- (5) selecting said transformed cells by their ability to grow in a selective medium lacking histidine.
A transformed Candida tropicalis host derived from C tropicalis SH-1 which is prototrophic as the result of transformation with the C. tropicalis HIS4 gene.
Dependencies are inferred from the claim text.