公开(公告)号：US4892819A

公开(公告)日：1990-01-09

申请号：US801523

申请日：1985-11-25

申请人： Lucinda G. Carr ,  Thomas D. Ingolia ,  Stephen W. Queener ,  Paul L. Skatrud

发明人： Lucinda G. Carr ,  Thomas D. Ingolia ,  Stephen W. Queener ,  Paul L. Skatrud

IPC分类号： C12N15/09 ,  C12N1/14 ,  C12N1/15 ,  C12N1/20 ,  C12N1/21 ,  C12N9/00 ,  C12N15/00 ,  C12N15/52 ,  C12N15/69 ,  C12N15/70 ,  C12N15/80 ,  C12N15/90 ,  C12P37/00 ,  C12R1/19 ,  C12R1/75 ,  C12R1/82

CPC分类号： C12N9/93 ,  C12N15/69 ,  C12N15/70 ,  C12N15/80 ,  C12N15/90

摘要： The present invention comprises novel DNA compounds that encode isopenicillin N synthetase. The invention also comprises methods, transformants, and polypeptides related to the novel DNA compounds. The novel isopenicillin N synthetase-encoding DNA, together with its associated transcription and translation activating sequence, was isolated from Penicillium chrysogenum. The isopenicillin N synthetase-encoding DNA can be used to construct novel E. coli expression vectors that drive expression of isopenicillin N synthetase in E. coli. The intact P. chrysogenum isopenicillin N synthetase-encoding DNA and associated transcription and translation activating sequence can also be used to construct expression vectors that drive expression of the isopenicillin N synthetase in P. chrysogenum and Cephalosporium acremonium. The transcription and translation activating sequence can be fused to a hygromycin phosphotransferase-encoding DNA segment and placed onto expression vectors that function in P. chrysogenum and C. acremonium. The transcription termination and mRNA polyadenylation signals of the P. chrysogenum isopenicillin N synthetase can be used to increase ultimate expression of a product encoded on a recombinant DNA vector.

摘要翻译： 本发明包括编码异青霉素N合成酶的新型DNA化合物。 本发明还包括与新型DNA化合物相关的方法，转化体和多肽。 编码新颖的异青霉素N合成酶的DNA，连同其相关的转录和翻译活化序列，从产黄青霉中分离出来。 编码异青霉素N合成酶的DNA可用于构建驱动大肠杆菌中异青霉素N合成酶表达的新型大肠杆菌表达载体。 完整的产黄青霉素异青霉素N合成酶编码DNA和相关的转录和翻译活化序列也可用于构建表达产物，其驱动产黄青霉和顶头孢霉的异青霉素N合成酶的表达。 转录和翻译激活序列可以与编码潮霉素磷酸转移酶的DNA片段融合，并置于表达载体中，该表达载体在产黄青霉和顶头孢霉中起作用。 产黄青霉素异青霉素N合成酶的转录终止和mRNA聚腺苷酸化信号可用于增加编码在重组DNA载体上的产物的最终表达。