公开(公告)号：US5965444A

公开(公告)日：1999-10-12

申请号：US880745

申请日：1997-06-23

申请人： Toshihiko Ashikari ,  Hiroto Kondo ,  Keiko Sakakibara ,  Hiroyuki Araki ,  Yasuji Oshima

发明人： Toshihiko Ashikari ,  Hiroto Kondo ,  Keiko Sakakibara ,  Hiroyuki Araki ,  Yasuji Oshima

IPC分类号： C12N9/34 ,  C12N15/65 ,  C12N15/81 ,  C12N15/90 ,  C12Q1/68 ,  C07H21/04

CPC分类号： C12N9/2428 ,  C12N15/65 ,  C12N15/81 ,  C12N15/905

摘要： A DNA construct wherein a DNA fragment which is recombinable in yeast chromosomal DNA is directly or indirectly linked at both ends of a DNA fragment which comprises a pair of R sensitive sequences oriented in the same direction and flaking both an R gene placed under the control of an inducible promoter and an expressible selective marker gene, which is a DNA construct designed with the R sensitive sequences non-symmetrically shortened, so that no functionable R sensitive sequence remains after the R sensitive sequence recombination has occurred by expression of the R gene and the selective marker has been removed. Since no functionable R sensitive sequence remains after removal of the selective marker, recombination does not occur again, and thus the same selective marker may be used for multiple insertions of foreign genes.

摘要翻译： 一种DNA构建体，其中在酵母染色体DNA中可重组的DNA片段在DNA片段的两端直接或间接连接，所述DNA片段包含一对沿相同方向取向的R敏感序列， 可诱导的启动子和可表达的选择性标记基因，其是用R敏感序列非对称缩短设计的DNA构建体，使得在通过R基因的表达发生R敏感序列重组后，不存在可操作的R敏感序列 选择性标记已被删除。 由于在去除选择性标记物之后不存在可操作的R敏感序列，因此重新不再次发生，因此相同的选择性标记可用于多次插入外源基因。

公开(公告)号：US4837147A

公开(公告)日：1989-06-06

申请号：US896176

申请日：1986-08-13

申请人： Yasuji Oshima ,  Hiroyuki Araki ,  Yoshinobu Kaneko

发明人： Yasuji Oshima ,  Hiroyuki Araki ,  Yoshinobu Kaneko

IPC分类号： C12N15/09 ,  C07K14/02 ,  C12N1/16 ,  C12N15/81 ,  C12P21/00 ,  C12R1/19 ,  C12R1/865

CPC分类号： C07K14/005 ,  C12N15/81 ,  C12N2730/10122

摘要： The present invention relates to a DNA fragment having a promoter activity of PHO81 gene regulating the production of phosphatase, and which is obtainable from Saccharomyces cerevisiae.; a DNA fragment bearing a structural gene downstream from the above PHO81 promoter; a transformant containing a DNA fragment bearing the above PHO81 promoter; a transformant containing a DNA fragment bearing the above PHO81 promoter and a structural gene downstream from the PHO81 promoter; and a process for obtaining a gene product which includes culturing a transformant containing a DNA fragment bearing the above PHO81 promoter and a structural gene located downstream therefrom in a suitable medium until the gene product is formed and recovering the gene product from the culture. Pharmacologically important proteinous materials may be efficiently produced with the use of the above-described novel and potent promoter obtained from yeast which is a eukaryotic microorganism.

摘要翻译： 本发明涉及具有调节磷酸酶生成的PHO81基因的启动子活性的DNA片段，其可从酿酒酵母获得。 具有上述PHO81启动子下游的结构基因的DNA片段; 含有携带上述PHO81启动子的DNA片段的转化体; 含有携带上述PHO81启动子的DNA片段和来自PHO81启动子下游的结构基因的转化体; 以及获得基因产物的方法，其包括培养含有携带上述PHO81启动子的DNA片段和位于其下游的结构基因的转化体在合适的培养基中，直到形成基因产物并从培养物中回收基因产物。 可以使用从真菌微生物酵母获得的上述新型有效的促进剂，有效地产生药理学上重要的蛋白质物质。