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发明公开
CN102304541A 一种含组成型启动子pPGK和G418抗性基因的酿酒酵母表达载体及其构建方法 无效 - 撤回

  • 专利标题： 一种含组成型启动子pPGK和G418抗性基因的酿酒酵母表达载体及其构建方法
  • 专利标题（英）： Saccharomyces cerevisiae expression carrier containing constitutive promoter pPGK and G418 resistance gene and construction method thereof
  • 申请号： CN201110235211.X
    申请日： 2011-08-17
  • 公开(公告)号： CN102304541A
    公开(公告)日： 2012-01-04
  • 发明人: 周长林 , 曹喜涛 , 陈凯 , 窦洁 , 王慧 , 李扬 , 奚涛
  • 申请人： 中国药科大学
  • 申请人地址： 江苏省南京市江宁区龙眠大道639号
  • 专利权人： 中国药科大学
  • 当前专利权人： 中国药科大学
  • 当前专利权人地址： 江苏省南京市江宁区龙眠大道639号
  • 主分类号： C12N15/81
  • IPC分类号： C12N15/81 ;  C12N15/66
一种含组成型启动子pPGK和G418抗性基因的酿酒酵母表达载体及其构建方法
摘要：
本发明公开了一种含组成型启动子pPGK和G418抗性基因的酿酒酵母表达载体及其构建方法，其目的是解决目前酿酒酵母表达载体多以营养缺陷为筛选标记，难以在工业酵母菌株进行基因工程改造过程中使用这一难题。其构建方法步骤如下：克隆KanMX基因，与pYES2.0载体连接，构建质粒pYES-Kan1。同时，以酿酒酵母染色体DNA为模板，克隆pPGK启动子基因，将其与pYES-Kan1质粒连接，将其原来的pGAL诱导型启动子替换为pPGK启动子，从而构建组成型酿酒酵母表达质粒pPGK-KanMX。本发明操作简单，构建的质粒无需对工业酵母菌株进行营养缺陷改造即可进行基因操作，大大简化了酵母基因工程改造过程。
摘要（英）：

The invention discloses a saccharomyces cerevisiae expression carrier containing a constitutive promoter pPGK gene and a G418 resistance gene and a construction method thereof, aiming at solving the difficulty that the existing saccharomyces cerevisiae expression carrier mainly takes nutritional deficiency as a screening mark, and is difficult to use in the genetic engineering modification process for an industrial yeast strain. The construction method comprises the following steps: cloning a KanMX gene; connecting with a pYES2.0 carrier; constructing a plasmid pYES-Kan1; and simultaneously cloning the promoter pPGK gene by taking saccharomyces cerevisiae chromosome DNA as a template, connecting the promoter pPGK gene with the plasmid pYES-Kan1, and replacing the original pGAL induced promoter with the promoter pPGK, thus the constitutive saccharomyces cerevisiae expression plasmid pPGK-KanMX is constructed. According to the invention, the operation is simple, the constructed plasmid is not required to perform nutrition deficiency modification on the industrial yeast strain, the gene operation can be carried out, and the modification process of the cerevisiae gene engineering is simplified greatly.

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