HANSENULA POLYMORPHA GENE AND STRAIN LACKING OF THE SAME GENE

摘要

PURPOSE: A Hansenula polymorpha gene and a strain lacking of the same gene are provided, thereby a foreign protein having decreased carboxy terminal digestion can be produced from the strain lacking of the Hansenula polymorpha gene. CONSTITUTION: A PEP4 gene having the nucleotide sequence of SEQ ID NO: 2 and encoding protease A derived from Hansenula polymorpha DL1(ATCC 26012). A recombinant vector pHPL is constructed by inserting Hansenula polymorpha LEU2 gene into a plasmid pHDP4 containing a gene PEP4 having the nucleotide sequence of SEQ ID NO: 2. A recombinant vector pHPUZ is constructed by inserting Hansenula polymorpha URA3 gene pop out cassette into a plasmid PHDP4 containing the gene PEP4 having the nucleotide sequence of SEQ ID NO:2. The Hansenula polymorpha UR2 and DL1 protease A mutant strains are produced by transforming with the recombinant vectors pHPL and pHPUZ, respectively. A foreign protein is expressed by independently inserting the foreign gene into the transformed mutant strains and independently culturing the strains in a methanol medium, wherein the foreign protein is hEGF.