公开(公告)号：US5258302A

公开(公告)日：1993-11-02

申请号：US560618

申请日：1990-07-30

Applicant: Thomas S. Vedvick ,  Michael E. Engel ,  Mary S. Urcan ,  Richard G. Buckholz ,  Jennifer A. Kinney

Inventor： Thomas S. Vedvick ,  Michael E. Engel ,  Mary S. Urcan ,  Richard G. Buckholz ,  Jennifer A. Kinney

IPC: A61K38/00 ,  C07K14/395 ,  C07K14/81 ,  C12N15/15 ,  C12N15/81 ,  C07K7/10 ,  C12N1/19 ,  C12P21/02

CPC classification number: C12N15/815 ,  C07K14/395 ,  C07K14/8117 ,  A61K38/00 ,  C07K2319/02

Abstract: 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.

Abstract translation: 生物活性抑肽酶（APR）分子，天然存在的，相对短的单链多肽，是通过生长甲基营养酵母转化体来制备的，所述转基因在其基因组中含有至少一个可操作地编码APR的DNA序列拷贝，与编码 酿酒酵母α交配因子前序序列（包括选自lys-arg-（glu-ala）x的加工序列，其中x是0-3范围内的整数），两者均在启动子 区域的甲基营养酵母的基因，允许所述DNA序列表达的条件和APR分子分泌到培养基中。 还公开了可用于本发明实践的新型DNA片段和新型重组酵母菌株。