公开(公告)号：US20130309772A1

公开(公告)日：2013-11-21

申请号：US13877225

申请日：2011-09-30

申请人： Keishi Sakaguchi ,  Rie Hamaguchi ,  Takanori Matsuda ,  Makoto Ito ,  Naoki Nagano ,  Masahiro Hayashi ,  Daisuke Honda ,  Yuji Okita ,  Shinichi Sugimoto

发明人： Keishi Sakaguchi ,  Rie Hamaguchi ,  Takanori Matsuda ,  Makoto Ito ,  Naoki Nagano ,  Masahiro Hayashi ,  Daisuke Honda ,  Yuji Okita ,  Shinichi Sugimoto

IPC分类号： C12N15/79 ,  C12N15/89

CPC分类号： C12N15/79 ,  C12N9/0071 ,  C12N9/0083 ,  C12N9/1029 ,  C12N15/113 ,  C12N15/52 ,  C12N15/895 ,  C12N2310/11 ,  C12N2310/14 ,  C12P7/6409 ,  C12P7/6427 ,  C12P7/6472 ,  C12Y114/19001 ,  C12Y114/19006 ,  C12Y203/01119

摘要： To provide a transformation method for producing a stramenopile organism having an improved unsaturated fatty acid production capability by disrupting a gene of the stramenopile organism or inhibiting the expression of the gene in a genetically engineering manner. [Solution] A method for transforming a stramenopile organism, which comprises disrupting a gene of the stramenopile organism or inhibiting the expression of the gene in a genetically engineering manner, and which is characterized in that the stramenopile organism is selected from Thraustochytrium aureum, Parietichytrium sarkarianum, Thraustochytrium roseum and Parietichytrium sp. and the gene to be disrupted or of which the expression is to be inhibited is a gene associated with the biosynthesis of a fatty acid.

摘要翻译： 提供通过以遗传工程方式破坏该层状生物体的基因或抑制该基因的表达来生产具有改善的不饱和脂肪酸生产能力的前烧石生物的转化方法。 [解决方案]一种转基因生物的方法，其包括以基因工程方式破坏所述地精生物体的基因或抑制所述基因的表达，其特征在于，所述前列腺生物体选自甲状腺破囊壶菌（Salisochytrium aureum），，鱼（Parietichytrium sarkarianum） ，玫瑰色胸，和Parietichytrium sp。 并且待破坏的基因或其表达被抑制的基因是与脂肪酸的生物合成相关的基因。

公开(公告)号：US20120322116A1

公开(公告)日：2012-12-20

申请号：US13497894

申请日：2010-09-24

申请人： Keishi Sakaguchi ,  Takanori Matsuda ,  Takumi Kobayashi ,  Makoto Ito ,  Naoki Nagano ,  Masahiro Hayashi ,  Daisuke Honda ,  Yosuke Taoka ,  Yuji Okita ,  Hitoshi Izumida ,  Shinichi Sugimoto

发明人： Keishi Sakaguchi ,  Takanori Matsuda ,  Takumi Kobayashi ,  Makoto Ito ,  Naoki Nagano ,  Masahiro Hayashi ,  Daisuke Honda ,  Yosuke Taoka ,  Yuji Okita ,  Hitoshi Izumida ,  Shinichi Sugimoto

IPC分类号： C12P7/64 ,  C12N15/79

CPC分类号： C12P7/6409 ,  C11C3/00 ,  C12N9/0071 ,  C12N9/0083 ,  C12N15/79 ,  C12N15/8201 ,  C12N15/8247 ,  C12P7/6427

摘要： Disclosed is a transformation method whereby an ability to produce a useful substance of a stramenopile can be improved. The method for transforming a stramenopile comprises transferring a foreign gene into the stramenopile which is a microorganism belonging to the class Labyrinthula, more specifically, to a genus Labyrinthula, Altornia, Aplanochytrium, Schizochytrium, Aurantiochytrium, Thraustochytrium, Ulkenia, etc. Said foreign gene, which is a gene relating to tolerance against an antibiotic, a colorimetric protein and/or a fatty acid desaturase (Δ5 desaturase gene, Δ12 desaturase gene and/or ω3 desaturase gene), is transferred by using the electroporation or gene-gun technique.

摘要翻译： 公开了能够提高产生有机物的能力的转化方法。 用于转化stramenopile的方法包括将外源基因转移到作为属于Labyrinthula类的微生物的地幔中，更具体地是属于Labyrinthula，Altornia，Aplanochytrium，Schizochytrium，Aurantiochytrium，Thraustochytrium，Ulkenia等。所述外源基因， 其是与抗生素，比色蛋白质和/或脂肪酸去饱和酶（＆Dgr。5去饱和酶基因，＆Dgr; 12去饱和酶基因和/或ω3去饱和酶基因）的耐受相关的基因通过使用电穿孔或基因 - 枪技术。

公开(公告)号：US09150891B2

公开(公告)日：2015-10-06

申请号：US13497894

申请日：2010-09-24

申请人： Keishi Sakaguchi ,  Takanori Matsuda ,  Takumi Kobayashi ,  Makoto Ito ,  Naoki Nagano ,  Masahiro Hayashi ,  Daisuke Honda ,  Yosuke Taoka ,  Yuji Okita ,  Hitoshi Izumida ,  Shinichi Sugimoto

发明人： Keishi Sakaguchi ,  Takanori Matsuda ,  Takumi Kobayashi ,  Makoto Ito ,  Naoki Nagano ,  Masahiro Hayashi ,  Daisuke Honda ,  Yosuke Taoka ,  Yuji Okita ,  Hitoshi Izumida ,  Shinichi Sugimoto

IPC分类号： C12P7/64 ,  C12P21/06 ,  C12N1/00 ,  C12N1/10 ,  C12N15/00 ,  C11C3/00 ,  C12N9/02 ,  C12N15/79 ,  C12N15/82

CPC分类号： C12P7/6409 ,  C11C3/00 ,  C12N9/0071 ,  C12N9/0083 ,  C12N15/79 ,  C12N15/8201 ,  C12N15/8247 ,  C12P7/6427

摘要： Disclosed is a transformation method whereby an ability to produce a useful substance of a stramenopile can be improved. The method for transforming a stramenopile comprises transferring a foreign gene into the stramenopile which is a microorganism belonging to the class Labyrinthula, more specifically, to a genus Labyrinthula, Altornia, Aplanochytrium, Schizochytrium, Aurantiochytrium, Thraustochytrium, Ulkenia, etc. Said foreign gene, which is a gene relating to tolerance against an antibiotic, a colorimetric protein and/or a fatty acid desaturase (Δ5 desaturase gene, Δ12 desaturase gene and/or ω3 desaturase gene), is transferred by using the electroporation or gene-gun technique.

摘要翻译： 公开了能够提高产生有机物的能力的转化方法。 用于转化stramenopile的方法包括将外源基因转移到作为属于Labyrinthula类的微生物的地幔中，更具体地是属于Labyrinthula，Altornia，Aplanochytrium，Schizochytrium，Aurantiochytrium，Thraustochytrium，Ulkenia等。所述外源基因， 其是与抗生素，比色蛋白质和/或脂肪酸去饱和酶（＆Dgr。5去饱和酶基因，＆Dgr; 12去饱和酶基因和/或ω3去饱和酶基因）的耐受相关的基因通过使用电穿孔或基因 - 枪技术。

公开(公告)号：US09062315B2

公开(公告)日：2015-06-23

申请号：US13877225

申请日：2011-09-30

申请人： Keishi Sakaguchi ,  Rie Hamaguchi ,  Takanori Matsuda ,  Makoto Ito ,  Naoki Nagano ,  Masahiro Hayashi ,  Daisuke Honda ,  Yuji Okita ,  Shinichi Sugimoto

发明人： Keishi Sakaguchi ,  Rie Hamaguchi ,  Takanori Matsuda ,  Makoto Ito ,  Naoki Nagano ,  Masahiro Hayashi ,  Daisuke Honda ,  Yuji Okita ,  Shinichi Sugimoto

IPC分类号： C12N15/79 ,  C12N15/89 ,  C12N9/02 ,  C12N9/10 ,  C12P7/64 ,  C12N15/52

CPC分类号： C12N15/79 ,  C12N9/0071 ,  C12N9/0083 ,  C12N9/1029 ,  C12N15/113 ,  C12N15/52 ,  C12N15/895 ,  C12N2310/11 ,  C12N2310/14 ,  C12P7/6409 ,  C12P7/6427 ,  C12P7/6472 ,  C12Y114/19001 ,  C12Y114/19006 ,  C12Y203/01119

摘要： To provide a transformation method for producing a stramenopile organism having an improved unsaturated fatty acid production capability by disrupting a gene of the stramenopile organism or inhibiting the expression of the gene in a genetically engineering manner. [Solution] A method for transforming a stramenopile organism, which comprises disrupting a gene of the stramenopile organism or inhibiting the expression of the gene in a genetically engineering manner, and which is characterized in that the stramenopile organism is selected from Thraustochytrium aureum, Parietichytrium sarkarianum, Thraustochytrium roseum and Parietichytrium sp. and the gene to be disrupted or of which the expression is to be inhibited is a gene associated with the biosynthesis of a fatty acid.

摘要翻译： 提供通过以遗传工程方式破坏该层状生物体的基因或抑制该基因的表达来生产具有改善的不饱和脂肪酸生产能力的前烧石生物的转化方法。 [解决方案]一种转基因生物的方法，其包括以基因工程方式破坏所述地精生物体的基因或抑制所述基因的表达，其特征在于，所述前列腺生物体选自甲状腺破囊壶菌（Salisochytrium aureum），，鱼（Parietichytrium sarkarianum） ，玫瑰色胸，和Parietichytrium sp。 并且待破坏的基因或其表达被抑制的基因是与脂肪酸的生物合成相关的基因。

公开(公告)号：US20120100550A1

公开(公告)日：2012-04-26

申请号：US13254794

申请日：2010-03-04

申请人： Naoki Nagano ,  Masahiro Hayashi ,  Keishi Sakaguchi ,  Makoto Ito ,  Yosuke Taoka ,  Yuji Okita

发明人： Naoki Nagano ,  Masahiro Hayashi ,  Keishi Sakaguchi ,  Makoto Ito ,  Yosuke Taoka ,  Yuji Okita

IPC分类号： C12Q1/68

CPC分类号： C12Q1/689

摘要： Disclosed is a method for determining the fatty acid synthesis pathway of a microorganism. Specifically disclosed is a method for determining the fatty acid synthesis pathway of a microorganism, which is characterized by producing degenerate primers for a fatty acid synthesis-related enzyme gene and determining the presence or absence of the fatty acid synthesis-related gene in the genome gene extracted from the microorganism using the degenerate primers. The sequences for degenerate primers for C20-elongase gene, which is a fatty acid synthesis-related enzyme gene, are ATHGARTWYTKBRTITTYGTICA (SEQ ID NO:1) and TARTRISWRTACATIADIAMRTG (SEQ ID NO:2), and the sequences for degenerate primers for ?4-desaturase gene are GGNCAYCAYCMITAYACNAA (SEQ ID NO:3) and TCDATYTGRTGIBWIARNCC (SEQ ID NO:4). The microorganism is one belonging to the class Labyrinthulea. Also specifically disclosed is a PCR primer set for use in the determination method. The method is useful as a method for screening a microorganism capable of synthesizing a fatty acid.

摘要翻译： 公开了一种测定微生物的脂肪酸合成途径的方法。 具体公开的是测定微生物的脂肪酸合成途径的方法，其特征在于制备脂肪酸合成相关酶基因的简并引物，并测定基因组基因中脂肪酸合成相关基因的存在或不存在 使用简并引物从微生物中提取。 作为脂肪酸合成相关酶基因的C20延伸酶基因的简并引物的序列是ATHGARTWYTKBRTITTYGTICA（SEQ ID NO：1）和TARTRISWRTACATIADIAMRTG（SEQ ID NO：2），并且用于Δ4的简并引物的序列 - 脱氢酶基因是GGNCAYCAYCMITAYACNAA（SEQ ID NO：3）和TCDATYTGRTGIBWIARNCC（SEQ ID NO：4）。 该微生物属于Labyrinthulea类。 还具体公开了用于确定方法的PCR引物组。 该方法可用作筛选能够合成脂肪酸的微生物的方法。