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1. 发明申请

WO1984001171A1 PLASMIDS WITH CONDITIONAL UNCONTROLLED REPLICATION BEHAVIOUR 审中-公开
标题翻译：具有条件不连续复制行为的PLASMIDS
公开(公告)号：WO1984001171A1
公开(公告)日：1984-03-29
申请号：PCT/DK1983000084
申请日：1983-09-09
申请人： A/S ALFRED BENZON , MOLIN, Soren , LARSEN, Jens, Erik, Love , LIGHT, Janice, Ann
发明人： A/S ALFRED BENZON
IPC分类号： C12N15/00
CPC分类号： C12N15/68 , C12N15/69
摘要： Plasmids useful as cloning vectors which carry a regulatable promoter which has been inserted in such a way that transcription from the promoter regulates replication so that increased transcription leads to a substantially increased or uncontrolled plasmid copy number when host microorganisms containing the plasmid are cultivated under conditions securing such increased transcription. The regulatable promoter is preferably lambda PR, the activity of which is controlled by a temperature-sensitive lambda cl repressor. To obtain gene products of the plasmids, host microorganisms to which such plasmids have been transformed are cultivated at about 30 C to secure a constant, low plasmid copy number during the seeding and multiplication stages of the microorganism, after which the temperature is shifted to about 36-42 C to obtain an uncontrolled plasmid copy number and an amplified amount of gene product. The gene product is then harvested from the culture.
摘要翻译：可用作携带可调节启动子的克隆载体的质粒，其以从启动子转录调节复制的方式插入，使得当含有质粒的宿主微生物的培养时，增加的转录导致显着增加或不受控制的质粒拷贝数，这种增加的转录。可调节的启动子优选为λPR，其活性由温度敏感的λ1c阻遏物控制。为了获得质粒的基因产物，在大约30℃培养这些质粒转化的宿主微生物，以在微生物的播种和繁殖阶段期间确保恒定的低质粒拷贝数，之后温度为转移到约36-42℃以获得不受控制的质粒拷贝数和扩增量的基因产物。然后从培养物中收获基因产物。

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