C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
195/1.15, 195/1.
C12N 15/81 (2006.01) C07K 14/02 (2006.01) C12N 1/19 (2006.01) C12N 15/00 (2006.01) C12N 15/63 (2006.01) C12N 15/90 (2006.01)
Patent
CA 1339209
Method for the site specific genomic modification of yeasts of the genus Pichia and novel DNA sequences useful therefor are provided. Pichia is transformed with a serially arranged linear DNA fragment comprising first and second insertable DNA fragments, which flank a marker gene. The insertable DNA sequences are homologous to defined portions of the Pichia genome and integrate by recombination at such defined sites. The resulting transformed organism contains the marker gene and any additional DNA sequences which are positioned between the insertable DNA fragments. In addition, the resulting transformed organism has either a disrupted or deleted sequence at the site of the genomic modification. Enhanced production of heterologous gene products is observed when using as the host for expression strains in which the alcohol oxidase gene has been disrupted. Upon disruption of the primary alcohol oxidase gene of Pichia, the existence of a second alcohol oxidase gene in Pichia was also discovered.
514838
Osler Hoskin & Harcourt Llp
Phillips Petroleum Company
Research Corporation Technologies Inc.
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