公开(公告)号：US20090098607A1

公开(公告)日：2009-04-16

申请号：US11873394

申请日：2007-10-16

申请人： Keisuke NAKAJOU ,  Yohei HOASHI ,  Toshiya KAI ,  Tadayuki UNO ,  Masaki OTAGIRI

发明人： Keisuke NAKAJOU ,  Yohei HOASHI ,  Toshiya KAI ,  Tadayuki UNO ,  Masaki OTAGIRI

IPC分类号： C12P21/04 ,  C12N15/00

CPC分类号： C07K14/805 ,  C12N15/815

摘要： The present invention provides a production method of a functional recombinant human hemoglobin, which comprises transforming a yeast of the genus Pichia with an expression vector containing a DNA encoding a human hemoglobin α-chain (Hbα), which is under regulation of a promoter functional in a yeast of the genus Pichia, and an expression vector containing a DNA encoding a human hemoglobin β-chain (Hbβ), which is under regulation of a promoter functional in a yeast of the genus Pichia, culturing the yeast in a medium, and recovering the human hemoglobin from the obtained culture. The present invention also provides an expression vector containing AOX1 promoter, DNA encoding Hbβ, terminator functional in a yeast of the genus Pichia, AOX1 promoter, DNA encoding Hbα and terminator functional in the yeast of the genus Pichia in this order from the upstream side in transcriptional direction.

摘要翻译： 本发明提供了一种功能性重组人血红蛋白的制备方法，其包括用含有编码人血红蛋白α链（Hbalpha）的DNA的表达载体转化毕赤酵母属的酵母，所述人类血红蛋白α-链（Hbalpha）是在 毕赤酵母属的酵母，以及含有编码人血红蛋白β链（Hbbeta）的DNA的表达载体，所述DNA是在毕赤酵母属酵母中起促进作用的启动子的调节下，在培养基中培养所述酵母，并回收 来自所得培养物的人血红蛋白。 本发明还提供了包含AOX1启动子，编码Hbbeta的DNA，毕赤酵母属的酵母中的终止子功能的终止子，AOX1启动子，编码Hbalpha的DNA和在毕赤酵母属的酵母中的终止子功能的表达载体， 转录方向。

公开(公告)号：US20080166769A1

公开(公告)日：2008-07-10

申请号：US11921329

申请日：2006-06-01

申请人： Tadayuki Uno ,  Yohei Hoashi ,  Naohisa Katayama ,  Toshiya Kai

发明人： Tadayuki Uno ,  Yohei Hoashi ,  Naohisa Katayama ,  Toshiya Kai

IPC分类号： C12N15/63 ,  C12N1/21

CPC分类号： C12N15/70 ,  C07K14/805 ,  C12N15/69 ,  C12P21/02

摘要： Provided is a high-copy-number, high-expression vector capable of producing a protein having satisfactory functions and activity of the same level as that of a natural form of the protein, in a large quantity and in a simple manner. Also provided is a vector including: (A) a target gene or a cloning site of the target gene, (B) a sequence element necessary for the high copying of the target gene, (C) a sequence element necessary for the expression of the target gene and a methionine aminopeptidase gene; a method of producing the vector; a transformant having the vector introduced therein; and a process for producing a protein using the transformant.

摘要翻译： 提供了能够以大量和简单的方式产生具有与天然形式的蛋白质相同水平的令人满意的功能和活性的蛋白质的高拷贝数高表达载体。 还提供了载体，其包括：（A）靶基因的靶基因或克隆位点，（B）靶基因的高拷贝所必需的序列元件，（C）表达所需靶序列所需的序列元件 靶基因和甲硫氨酸氨基肽酶基因; 生成载体的方法; 其中引入载体的转化体; 以及使用该转化体生产蛋白质的方法。