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公开(公告)号：US20110262998A1
公开(公告)日：2011-10-27
申请号：US13125328
申请日：2009-10-21
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。
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公开(公告)号：US08748132B2
公开(公告)日：2014-06-10
申请号：US13125328
申请日：2009-10-21
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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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
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。
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