专利快速检索-快速检索全球专利，免费商用专利数据库-IPRDB

12. 发明授权

US4643969A Novel cloning vehicles for polypeptide expression in microbial hosts 失效
标题翻译：用于微生物宿主中多肽表达的新型克隆载体
公开(公告)号：US4643969A
公开(公告)日：1987-02-17
申请号：US494040
申请日：1983-07-25
申请人： Masayori Inouye , Yoshihiro Masui
发明人： Masayori Inouye , Yoshihiro Masui
IPC分类号： C12N15/62 , C12N15/70 , C12P21/00 , C12N1/00 , C12N15/00
CPC分类号： C12N15/62 , C12N15/70 , Y10S930/30
摘要： Methods and compositions are provided for regulated expression of polypeptides in transformed bacterial hosts. A novel class of plasmid cloning vehicles includes a DNA sequence coding for the desired polypeptide (on an insertion site therefor) linked for transcriptional expression in reading phase with four functional fragments derived from the lipoprotein gene of E. coli. The plasmids further include a DNA sequence coding for a specific segment of the E. coli lac promoter-operator, which is positioned in the proper orientation for transcriptional expression of the desired polypeptide, as well as a separate functional E. coli lacI gene coding for the associated repressor molecule which can interact with the lac promoter-operator to prevent transcription therefrom. Expression of the desired polypeptide is under the control of both the constitutive promoter and the inducible promoter, although transcription from either promotor is normally blocked by the repressor molecule. However, the repressor can be selectively inactivated by means of an inducer molecule to permit transcriptional expression of the desired polypeptide from both promoter. The methods utilize such plasmids to introduce genetic capability into micro-organisms for the production of proteins, such as medically or commerically useful hormones, enzymes, immunogenic proteins, or intermediates therefor, but only in the presence of an appropriate inducer.
摘要翻译：提供了用于在转化的细菌宿主中调节多肽表达的方法和组合物。一类新颖的质粒克隆载体包括编码与在大肠杆菌的脂蛋白基因中衍生的四个功能片段相连的用于在阅读阶段转录表达的期望多肽（在其插入位点上）的DNA序列。质粒还包括编码大肠杆菌lac启动子 - 操纵子的特定区段的DNA序列，其定位于所需多肽的转录表达的正确取向，以及编码所述多肽的单独的功能性大肠杆菌lacI基因可与lac启动子 - 操纵子相互作用以阻止转录的相关阻遏物分子。所需多肽的表达受组成型启动子和诱导型启动子的控制，尽管来自启动子的转录通常被阻遏物分子阻断。然而，阻遏物可以通过诱导剂分子选择性地失活以允许来自两个启动子的所需多肽的转录表达。所述方法利用这种质粒将遗传能力引入微生物以产生蛋白质，例如医学或商业上有用的激素，酶，免疫原性蛋白质或其中间体，但仅在适当的诱导剂存在下。

IPRDB

热门服务

关于我们

友情链接

联系方式