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1.发明申请公开(公告)号：US20080311658A1
公开(公告)日：2008-12-18
申请号：US12052658
申请日：2008-03-20
申请人： John Shanklin , Tam Nguyen
发明人： John Shanklin , Tam Nguyen
CPC分类号： C12N15/113 , C12N15/111 , C12N15/8218 , C12N15/8247 , C12N2310/11 , C12N2310/14 , C12N2310/3519 , C12N2310/53
摘要： A nucleotide construct comprising a nucleotide sequence that forms a stem and a loop, wherein the loop comprises a nucleotide sequence that modulates expression of a target, wherein the stem comprises a nucleotide sequence that modulates expression of a target, and wherein the target modulated by the nucleotide sequence in the loop and the target modulated by the nucleotide sequence in the stem may be the same or different. Vectors, methods of regulating target expression, methods of providing a cell, and methods of treating conditions comprising the nucleotide sequence are also disclosed.
摘要翻译： 包含形成茎和环的核苷酸序列的核苷酸构建体，其中所述环包含调节靶表达的核苷酸序列，其中所述茎包含调节靶表达的核苷酸序列，并且其中由 环中的核苷酸序列和由茎中的核苷酸序列调节的靶可以相同或不同。 还公开了载体，调节靶标表达的方法，提供细胞的方法和治疗包含该核苷酸序列的病症的方法。
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公开(公告)号：US06974893B2
公开(公告)日：2005-12-13
申请号：US10185578
申请日：2002-06-28
申请人： John Shanklin , Edward J. Whittle
发明人： John Shanklin , Edward J. Whittle
CPC分类号： C12N9/0073 , C12N15/8247
摘要： The present invention relates to oleate hydroxylase genes, proteins, and methods of their use. The present invention also relates to methods of using the oleate hydroxylase genes and proteins, including in their expression in transgenic organisms and in the production of hydroxylated fatty acids.
摘要翻译： 本发明涉及油酸羟化酶基因，蛋白质及其使用方法。 本发明还涉及使用油酸羟化酶基因和蛋白质的方法，包括在转基因生物中的表达和羟基化脂肪酸的生产。
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公开(公告)号：US20050005325A1
公开(公告)日：2005-01-06
申请号：US10822370
申请日：2004-04-12
申请人： John Shanklin , Edward Whittle
发明人： John Shanklin , Edward Whittle
CPC分类号： C12N15/8247 , C12N9/0083
摘要： The present invention relates to methods for producing fatty acid desaturase mutants having a substantially increased activity towards substrates with fewer than 18 carbon atom chains relative to an unmutagenized precursor desaturase having an 18 carbon chain length specificity, the sequences encoding the desaturases and to the desaturases that are produced by the methods. The present invention further relates to a method for altering a function of a protein, including a fatty acid desaturase, through directed mutagenesis involving identifying candidate amino acid residues, producing a library of mutants of the protein by simultaneously randomizing all amino acid candidates, and selecting for mutants which exhibit the desired alteration of function. Candidate amino acids are identified by a combination of methods. Enzymatic, binding, structural and other functions of proteins can be altered by the method.
摘要翻译： 本发明涉及生产脂肪酸去饱和酶突变体的方法，其具有相对于具有18个碳链长特异性的未成熟前体去饱和酶相对于具有少于18个碳原子链的底物具有显着增加的活性，编码去饱和酶的序列和去饱和酶， 是通过这些方法生产的。 本发明还涉及通过定向诱变来改变包括脂肪酸去饱和酶在内的蛋白质的功能的方法，所述定向诱变涉及鉴定候选氨基酸残基，通过同时随机化所有氨基酸候选物产生蛋白质突变体文库，并选择 对于表现出所需功能改变的突变体。 候选氨基酸通过方法的组合来鉴定。 可通过该方法改变蛋白质的酶，结合，结构和其他功能。
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公开(公告)号：US6100091A
公开(公告)日：2000-08-08
申请号：US276295
申请日：1999-03-25
IPC分类号： C12N15/09 , C12N1/15 , C12N1/21 , C12N5/10 , C12N9/02 , C12N15/00 , C12N15/11 , C12N15/29 , C12N15/82 , C07H21/04 , C12N5/14
CPC分类号： C12N9/0083 , C12N15/8247 , C07K2319/00
摘要： Disclosed is a methods for modifying the chain length and double bond positional specificities of a soluble plant fatty acid desaturase. More specifically, the method involves modifying amino acid contact residues in the substrate binding channel of the soluble fatty acid desaturase which contact the fatty acid. Specifically disclosed is the modification of an acyl-ACP desaturase. Amino acid contact residues which lie within the substrate binding channel are identified, and subsequently replaced with different residues to effect the modification of activity.
摘要翻译： 公开了用于改变可溶性植物脂肪酸去饱和酶的链长和双键位置特异性的方法。 更具体地，该方法包括修饰与脂肪酸接触的可溶性脂肪酸去饱和酶的底物结合通道中的氨基酸接触残基。 具体公开的是酰基-ACP去饱和酶的修饰。 鉴定位于底物结合通道内的氨基酸接触残基，随后用不同的残基替换以影响活性的修饰。
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5.发明授权公开(公告)号：US08796442B2
公开(公告)日：2014-08-05
申请号：US12052658
申请日：2008-03-20
申请人： John Shanklin , Tam Nguyen
发明人： John Shanklin , Tam Nguyen
IPC分类号： C07H21/04
CPC分类号： C12N15/113 , C12N15/111 , C12N15/8218 , C12N15/8247 , C12N2310/11 , C12N2310/14 , C12N2310/3519 , C12N2310/53
摘要： A nucleotide construct comprising a nucleotide sequence that forms a stem and a loop, wherein the loop comprises a nucleotide sequence that modulates expression of a target, wherein the stem comprises a nucleotide sequence that modulates expression of a target, and wherein the target modulated by the nucleotide sequence in the loop and the target modulated by the nucleotide sequence in the stem may be the same or different. Vectors, methods of regulating target expression, methods of providing a cell, and methods of treating conditions comprising the nucleotide sequence are also disclosed.
摘要翻译： 包含形成茎和环的核苷酸序列的核苷酸构建体，其中所述环包含调节靶表达的核苷酸序列，其中所述茎包含调节靶表达的核苷酸序列，并且其中由 环中的核苷酸序列和由茎中的核苷酸序列调节的靶可以相同或不同。 还公开了载体，调节靶标表达的方法，提供细胞的方法和治疗包含该核苷酸序列的病症的方法。
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公开(公告)号：US20100281573A1
公开(公告)日：2010-11-04
申请号：US12686975
申请日：2010-01-13
申请人： Ingo H. Heilmann , John Shanklin
发明人： Ingo H. Heilmann , John Shanklin
IPC分类号： C12N15/82
CPC分类号： C12N9/0083 , C07H21/04 , C12N15/8247
摘要： The present invention relates to enzymes involved in lipid metabolism. In particular, the present invention provides coding sequences for Arabidopsis Desaturases (ADS), the encoded ADS polypeptides, and methods for using the sequences and encoded polypeptides, where such methods include decreasing and increasing saturated fatty acid content in plant seed oils.
摘要翻译： 本发明涉及参与脂质代谢的酶。 特别地，本发明提供了拟南芥去饱和酶（ADS）的编码序列，编码的ADS多肽，以及使用序列和编码多肽的方法，其中这些方法包括减少和增加植物种子油中的饱和脂肪酸含量。
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公开(公告)号：US5888790A
公开(公告)日：1999-03-30
申请号：US853979
申请日：1997-05-09
IPC分类号： C12N15/09 , C12N1/15 , C12N1/21 , C12N5/10 , C12N9/02 , C12N15/00 , C12N15/11 , C12N15/29 , C12N15/82
CPC分类号： C12N9/0083 , C12N15/8247 , C07K2319/00
摘要： Disclosed is a method for modifying the chain length and double bond positional specificities of a soluble plant fatty acid desaturase. More specifically, the method involves modifying amino acid contact residues in the substrate binding channel of the soluble fatty acid desaturase which contact the fatty acid. Specifically disclosed is the modification of an acyl-ACP desaturase. Amino acid contact residues which lie within the substrate binding channel are identified, and subsequently replaced with different residues to effect the modification of activity.
摘要翻译： 公开了一种改变可溶性植物脂肪酸去饱和酶的链长和双键位置特异性的方法。 更具体地，该方法包括修饰与脂肪酸接触的可溶性脂肪酸去饱和酶的底物结合通道中的氨基酸接触残基。 具体公开的是酰基-ACP去饱和酶的修饰。 鉴定位于底物结合通道内的氨基酸接触残基，随后用不同的残基替换以影响活性的修饰。
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公开(公告)号：US20140230091A1
公开(公告)日：2014-08-14
申请号：US14130039
申请日：2012-06-28
申请人： John Shanklin , Carl Andre
发明人： John Shanklin , Carl Andre
IPC分类号： C12N15/82
CPC分类号： C12N15/8247 , C12N9/93
摘要： Methods and means are provided to increase the oil content of plants, particularly oleaginous plants by preventing feedback inhibition by 18:1-Coenzyme A or 18:1-Acyl Carrier Protein of the acetyl CoA-carboxylase enzyme in cells of these plants in various manners.
摘要翻译： 提供方法和手段，通过以各种方式防止这些植物细胞中18：1-辅酶A或18：1-酰基载体蛋白的乙酰辅酶A羧化酶的反馈抑制来增加植物，特别是含油植物的油含量 。
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公开(公告)号：US07323335B2
公开(公告)日：2008-01-29
申请号：US10822370
申请日：2004-04-12
申请人： John Shanklin , Edward J. Whittle
发明人： John Shanklin , Edward J. Whittle
CPC分类号： C12N15/8247 , C12N9/0083
摘要： The present invention relates to methods for producing fatty acid desaturase mutants having a substantially increased activity towards substrates with fewer than 18 carbon atom chains relative to an unmutagenized precursor desaturase having an 18 carbon chain length specificity, the sequences encoding the desaturases and to the desaturases that are produced by the methods. The present invention further relates to a method for altering a function of a protein, including a fatty acid desaturase, through directed mutagenesis involving identifying candidate amino acid residues, producing a library of mutants of the protein by simultaneously randomizing all amino acid candidates, and selecting for mutants which exhibit the desired alteration of function. Candidate amino acids are identified by a combination of methods. Enzymatic, binding, structural and other functions of proteins can be altered by the method.
摘要翻译： 本发明涉及生产脂肪酸去饱和酶突变体的方法，其具有相对于具有18个碳链长特异性的未成熟前体去饱和酶相对于具有少于18个碳原子链的底物具有显着增加的活性，编码去饱和酶的序列和去饱和酶， 是通过这些方法生产的。 本发明还涉及通过定向诱变来改变包括脂肪酸去饱和酶在内的蛋白质的功能的方法，所述定向诱变涉及鉴定候选氨基酸残基，通过同时随机化所有氨基酸候选物产生蛋白质突变体文库，并选择 对于表现出所需功能改变的突变体。 候选氨基酸通过方法的组合来鉴定。 可通过该方法改变蛋白质的酶，结合，结构和其他功能。
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公开(公告)号：US06686186B2
公开(公告)日：2004-02-03
申请号：US09988929
申请日：2001-12-03
申请人： John Shanklin , Edgar B. Cahoon
发明人： John Shanklin , Edgar B. Cahoon
IPC分类号： C12N902
CPC分类号： C12N9/0083 , C12N15/8247
摘要： The present invention relates to a method for producing mutants of a fatty acid desaturase having a substantially increased activity towards fatty acid substrates with chains containing fewer than 18 carbons relative to an unmutagenized precursor desaturase having an 18 carbon atom chain length substrate specificity. The method involves inducing one or more mutations in the nucleic acid sequence encoding the precursor desaturase, transforming the mutated sequence into an unsaturated fatty acid auxotroph cell such as MH13 E. coli, culturing the cells in the absence of supplemental unsaturated fatty acids, thereby selecting for recipient cells which have received and which express a mutant fatty acid desaturase with an elevated specificity for fatty acid substrates having chain lengths of less than 18 carbon atoms. A variety of mutants having 16 or fewer carbon atom chain length substrate specificities are produced by this method. Mutant desaturases produced by this method can be introduced via expression vectors into prokaryotic and eukaryotic cells and can also be used in the production of transgenic plants which may be used to produce specific fatty acid products.
摘要翻译： 本发明涉及一种生产脂肪酸去饱和酶突变体的方法，该突变体相对于具有18个碳原子链长基底特异性的未成熟前体去饱和酶，具有对具有少于18个碳的链的脂肪酸底物具有显着增加的活性。 该方法包括在编码前体去饱和酶的核酸序列中诱导一个或多个突变，将突变的序列转化为不饱和脂肪酸营养缺陷型细胞如MH13大肠杆菌，在不存在补充不饱和脂肪酸的情况下培养细胞，从而选择 对于接受并且表达具有链长度小于18个碳原子的脂肪酸底物具有特异性提高的突变脂肪酸去饱和酶的受体细胞。 通过该方法制备具有16个或更少碳原子链长基底特异性的各种突变体。 通过该方法产生的突变型去饱和酶可以通过表达载体导入原核和真核细胞中，并且还可用于生产可用于产生特定脂肪酸产物的转基因植物。
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