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1. 发明申请

US20110262998A1 PROCESS FOR PREPARING INCLUSION BODY-FORMING PROTEIN 有权
标题翻译：制备包裹体形成蛋白的方法
公开(公告)号：US20110262998A1
公开(公告)日：2011-10-27
申请号：US13125328
申请日：2009-10-21
申请人： Hiroshi Nakatake , Akihiro Meta , Kiyotaka Suenaga , Masaki Hirashima , Hiroaki Maeda
发明人： Hiroshi Nakatake , Akihiro Meta , Kiyotaka Suenaga , Masaki Hirashima , Hiroaki Maeda
IPC分类号： C12N9/52 , C12N15/63 , C12N1/21 , C12N15/57
CPC分类号： C12N9/6491 , C07K2319/02 , C12P21/02
摘要： A process for preparing an inclusion body-forming protein is provided. A nucleic acid fragment consisting of a nucleotide sequence coding for a modified alkaline phosphatase signal peptide (modified APSP) where leucine at the 13th position in the amino acid sequence shown in SEQ ID NO: 1 is substituted with proline and/or alanine at the 21st position is substituted with the other amino acid, downstream of which nucleotide sequence is bound a nucleotide sequence of a gene of a protein of interest, and a process for preparing an inclusion body-forming protein consisting of the following steps: (1) preparing an expression vector in which a nucleic acid fragment is incorporated consisting of a nucleotide sequence coding for a modified signal peptide downstream of which is bound a nucleotide sequence of a gene of a protein of interest, (2) preparing a host cell transformed with the expression vector that produces an inclusion body-forming protein, and (3) purifying the inclusion body-forming protein from culture obtained by culturing the host cell that produces the inclusion body-forming protein. The above other amino acid is selected from the group consisting of aspartic acid, glutamic acid, lysine, histidine, phenylalanine and tyrosine.
摘要翻译：提供了一种制备包涵体形成蛋白的方法。由编码改变的碱性磷酸酶信号肽（修饰的APSP）的核苷酸序列组成的核酸片段，其中在第21位被脯氨酸和/或丙氨酸取代在SEQ ID NO：1所示的氨基酸序列中的第13位的亮氨酸位置被其他氨基酸取代，其中核苷酸序列的下游与感兴趣蛋白质的基因的核苷酸序列结合，以及由以下步骤组成的包涵体形成蛋白的方法：（1）制备表达载体，其中掺入核酸片段由编码下游的修饰信号肽的核苷酸序列组成，所述核苷酸序列与感兴趣蛋白质的基因的核苷酸序列结合，（2）制备用表达载体转化的宿主细胞产生包涵体形成蛋白质，（3）通过培养生成的宿主细胞获得的培养物纯化包涵体形成蛋白质是包涵体形成蛋白。上述其它氨基酸选自天冬氨酸，谷氨酸，赖氨酸，组氨酸，苯丙氨酸和酪氨酸。

2. 发明申请

US20080064061A1 Yeast Promoter 审中-公开
标题翻译：酵母启动子
公开(公告)号：US20080064061A1
公开(公告)日：2008-03-13
申请号：US11659492
申请日：2005-08-02
申请人： Akihiro Meta , Yo Nakahara , Hirofumi Higuchi
发明人： Akihiro Meta , Yo Nakahara , Hirofumi Higuchi
IPC分类号： C12P21/04 , C12N1/00 , C12N15/00 , C12N15/11
CPC分类号： C12N15/81
摘要： The present invention provides a DNA fragment exhibiting a potent promoter activity when used in the yeast. The present invention relates to a DNA fragment possessing a transcription promoter activity of the yeast, said DNA fragment having a whole of the DNA sequence shown by SEQ ID NO: 1 or a portion of said DNA sequence inclusive of its 3′-end, or having a DNA sequence which is hybridizable to a sequence complementary to a whole of the DNA sequence shown by SEQ ID NO: 1 or to a portion of said DNA sequence inclusive of its 3′-end and maintains the transcription promoter activity of said whole or portion of the DNA sequence shown by SEQ ID NO: 1; an expression plasmid containing a recombinant DNA fragment comprising said DNA fragment and a gene coding for a heterologous protein positioned downstream of said DNA sequence; a host cell transformed with said recombinant DNA fragment or said expression plasmid; and a method for preparing a heterologous protein which comprises culturing said host cell and recovering the heterologous protein from the culture. According to the present invention, a potent promoter usable in a heterologous gene expression system with the yeast as a host is provided.
摘要翻译：本发明提供了当在酵母中使用时显示出有效启动子活性的DNA片段。本发明涉及具有酵母的转录启动子活性的DNA片段，所述DNA片段具有SEQ ID NO：1所示的整个DNA序列或所述DNA序列的一部分（包括其3'末端），或具有可与SEQ ID NO：1所示的整个DNA序列互补的序列杂交的DNA序列或包含其3'末端的所述DNA序列的一部分，并保持所述整体的转录启动子活性或 SEQ ID NO：1所示的DNA序列的部分; 含有包含所述DNA片段的重组DNA片段和编码位于所述DNA序列下游的异源蛋白质的基因的表达质粒; 用所述重组DNA片段或所述表达质粒转化的宿主细胞; 以及制备异源蛋白质的方法，其包括培养所述宿主细胞并从培养物中回收异源蛋白质。根据本发明，提供了可用于以酵母作为宿主的异源基因表达系统中的有效启动子。

3. 发明授权

US08748132B2 Process for preparing inclusion body-forming protein 有权
标题翻译：制备包涵体形成蛋白的方法
公开(公告)号：US08748132B2
公开(公告)日：2014-06-10
申请号：US13125328
申请日：2009-10-21
申请人： Hiroshi Nakatake , Akihiro Meta , Kiyotaka Suenaga , Masaki Hirashima , Hiroaki Maeda
发明人： Hiroshi Nakatake , Akihiro Meta , Kiyotaka Suenaga , Masaki Hirashima , Hiroaki Maeda
IPC分类号： C12P21/04
CPC分类号： C12N9/6491 , C07K2319/02 , C12P21/02
摘要： A process for preparing an inclusion body-forming protein is provided. A nucleic acid fragment having a nucleotide sequence coding for a modified alkaline phosphatase signal peptide (modified APSP) where leucine at the 13th position in the amino acid sequence shown in SEQ ID NO: 1 is substituted with proline and/or alanine at the 21st position is substituted with the other amino acid, downstream of which nucleotide sequence is bound a nucleotide sequence of a gene of a protein of interest is also provided.
摘要翻译：提供了一种制备包涵体形成蛋白的方法。具有编码改变的碱性磷酸酶信号肽（修饰的APSP）的核苷酸序列的核酸片段，其中在SEQ ID NO：1所示的氨基酸序列中的第13位的亮氨酸被第21位的脯氨酸和/或丙氨酸取代还提供了另一个氨基酸，其下游的核苷酸序列与目的蛋白质的基因的核苷酸序列结合。

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