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公开(公告)号：US20130084641A1
公开(公告)日：2013-04-04
申请号：US13708479
申请日：2012-12-07
申请人： Shib Sankar Basu , Kristern Kamerath Dang , Kasimalai Azhakanandam , Thomas Z. McNeill , Sheng Guo
发明人： Shib Sankar Basu , Kristern Kamerath Dang , Kasimalai Azhakanandam , Thomas Z. McNeill , Sheng Guo
IPC分类号： C12N15/10
CPC分类号： C12N15/1089 , C12N15/1034
摘要： The present invention provides a method of selecting a mRNA for production of a polypeptide of interest comprising: a) producing an array of individual mRNA sequences comprising different nucleotide sequences encoding the polypeptide of interest; b) determining one or more or two or more of the following parameters for each individual mRNA sequence of (a): (i) minimum free energy (MFE) RNA secondary structure; (ii) ensemble free energy (EFE); (iii) frequency of the minimum free energy (FMFE) RNA secondary structure in a thermodynamic ensemble; and (iv) ensemble diversity (ED); c) ranking the individual mRNA sequences of the array according the parameters determined in step (b); and d) selecting a mRNA sequence from the ranked array of step (c), wherein the selected mRNA produces the polypeptide of interest. The present invention further provides a method of selecting a mRNA for enhanced and reduced production of a polypeptide of interest.
摘要翻译： 本发明提供了选择产生感兴趣多肽的mRNA的方法，包括：a）产生包含编码目的多肽的不同核苷酸序列的单个mRNA序列的阵列; b）确定（a）的每个单独mRNA序列的一个或多个或两个或更多个以下参数：（i）最小自由能（MFE）RNA二级结构; （ii）合奏自由能（EFE）; （iii）热力学集合中最小自由能（FMFE）RNA二级结构的频率; 和（iv）集体多样性（ED）; c）根据步骤（b）中确定的参数对阵列的各个mRNA序列进行排序; 和d）从步骤（c）的排列阵列中选择mRNA序列，其中所选择的mRNA产生感兴趣的多肽。 本发明还提供了选择用于增强和减少感兴趣的多肽的产生的mRNA的方法。
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2.发明申请METHODS AND COMPOSITIONS FOR ENHANCED GENE EXPRESSION THROUGH INTRON FREE ENERGY REDUCTION 审中-公开通过免疫能量减少增强基因表达的方法和组合公开(公告)号：US20120096583A1
公开(公告)日：2012-04-19
申请号：US13233282
申请日：2011-09-15
CPC分类号： C12N15/8257 , C12N15/67 , C12N15/8216 , G16B20/00 , G16B25/00 , G16B30/00
摘要： The present invention provides a method for selecting an intron for enhanced expression of a nucleotide sequence encoding a polypeptide, comprising: a) determining a mean free energy value per base pair of one or more introns; b) determining a mean IMEter score of the one or more introns of (a); and c) selecting an intron having a mean free energy value per base pair of below about −0.268 kcal/mol/bp and a mean IMEter score of at least about −0.034/bp, thereby selecting an intron for enhanced expression of a nucleotide sequence encoding the polypeptide, wherein expression of a nucleotide sequence operably associated with the selected intron is enhanced as compared to expression of a nucleotide sequence not operably associated with an intron having a mean free energy value per base pair of below about −0.268 kcal/mol/bp and a mean IMEter score of at least about −0.034/bp.
摘要翻译： 本发明提供了选择用于增强编码多肽的核苷酸序列表达的内含子的方法，其包括：a）确定每个碱基对的一个或多个内含子的平均自由能值; b）确定（a）的一个或多个内含子的平均IMEter得分; 和c）选择具有低于约-0.268kcal / mol / bp的每碱基对的平均自由能值的内含子和至少约-0.034 / bp的平均IMEter得分，从而选择用于增强核苷酸序列表达的内含子 编码多肽，其中与所选内含子可操作地相关联的核苷酸序列的表达与不具有与每个碱基对的平均自由能值低于约-0.268千卡/摩尔/升的内含子不可操作地相关联的核苷酸序列的表达增强， bp和至少约-0.034 / bp的平均IMEter得分。
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公开(公告)号：US08642839B2
公开(公告)日：2014-02-04
申请号：US12813681
申请日：2010-06-11
IPC分类号： A01H5/10 , C07K14/415 , G01N33/564
CPC分类号： C12N15/8241 , C12N15/8216
摘要： Compositions and methods for transiently expressing proteins in a plant are provided. The compositions comprise plants, seeds, plant tissues, and plant parts expressing a protein, wherein the protein is expressed transiently and the transient expression of the protein can be used as a predictive model of how said protein will be expressed in stable transgenic plants in regards to qualitative and quantitative data. The predictive model may be used but is not limited to: promoter evaluation, evaluation of expression cassette construction for best performance (e.g. addition of enhancers or gene silencing suppressors), evaluation of best ways to express heterologous genes (e.g. point mutations, targeting), fast evaluation of endogenous gene knockout, evaluation of protein expression levels, cellular targeting, tissue targeting, transcriptional enhancers, translational enhancer protein toxicity and metabolic profiling. Further provided are methods of use.
摘要翻译： 提供了在植物中瞬时表达蛋白质的组合物和方法。 组合物包含植物，种子，植物组织和表达蛋白质的植物部分，其中蛋白质被瞬时表达，并且蛋白质的瞬时表达可以用作如何在稳定的转基因植物中表达所述蛋白质的预测模型 定性和定量数据。 可以使用预测模型，但不限于：启动子评估，用于最佳性能的表达盒构建的评估（例如添加增强子或基因沉默抑制基因），评估表达异源基因的最佳方式（例如点突变，靶向）， 内源基因敲除的快速评价，蛋白质表达水平的评价，细胞靶向，组织靶向，转录增强子，翻译增强子蛋白毒性和代谢分析。 进一步提供使用的方法。
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4.发明申请STACKING OF TRANSLATIONAL ENHANCER ELEMENTS TO INCREASE POLYPEPTIDE EXPRESSION IN PLANTS 审中-公开翻译增强元件的堆叠，以增加植物中的多肽表达公开(公告)号：US20120185969A1
公开(公告)日：2012-07-19
申请号：US13393846
申请日：2010-09-02
CPC分类号： C12N15/8216
摘要： Compositions and methods for increasing expression of a polypeptide of interest in a plant or plant part thereof are provided. Compositions of the invention are polynucleotide constructs comprising (a) at least one translational enhancer element derived from a virus tandemly stacked with at least one translational enhancer element derived from a cellular gene, and (b) an operably linked polynucleotide encoding a polypeptide of interest. Expression cassettes, vectors, and transgenic plants and plant parts comprising these polynucleotide constructs are also provided. Methods for increasing expression of a polypeptide of interest in a plant or plant part thereof utilizing the polynucleotide constructs and expression cassettes of the invention are also provided.
摘要翻译： 提供了用于增加植物或植物部分中感兴趣的多肽的表达的组合物和方法。 本发明的组合物是多核苷酸构建体，其包含（a）至少一种衍生自与来自细胞基因的至少一种翻译增强子串联堆叠的病毒的翻译增强子元件，和（b）编码目的多肽的可操作地连接的多核苷酸。 还提供了表达盒，载体和包含这些多核苷酸构建体的转基因植物和植物部分。 还提供了利用本发明的多核苷酸构建体和表达盒在其植物或植物部分中增加目的多肽表达的方法。
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5.发明授权公开(公告)号：US09068191B2
公开(公告)日：2015-06-30
申请号：US13326786
申请日：2011-12-15
申请人： Kasimalai Azhakanandam , Pei Su
发明人： Kasimalai Azhakanandam , Pei Su
CPC分类号： C12N15/8205 , C12N15/8202 , C12N15/8207 , C12N15/8216 , C12N15/8257
摘要： This invention provides methods of transiently expressing a nucleotide sequence in one or more cells of a soybean plant, comprising: a) abrading the surface of a soybean plant and/or intact part thereof; and either (b1) immersing the abraded plant and/or abraded intact part thereof in a solution comprising Agrobacterium cells and a surfactant, wherein the Agrobacterium cells comprise the nucleotide sequence to be transiently expressed; (b2) applying a vacuum to the immersed soybean plant and/or intact part thereof of step (b1); and (b3) releasing the vacuum, thereby introducing the solution into the plant and/or intact part thereof; or (c) contacting the abraded plant and/or abraded intact part thereof with a solution comprising Agrobacterium cells and a surfactant, wherein the Agrobacterium cells comprise the nucleotide sequence to be transiently expressed, whereby the nucleotide sequence is transiently expressed in one or more cells of the plant.
摘要翻译： 本发明提供了在大豆植物的一个或多个细胞中瞬时表达核苷酸序列的方法，包括：a）研磨大豆植物和/或其完整部分的表面; （b1）将磨损的植物和/或磨损的整体部分浸入包含农杆菌细胞和表面活性剂的溶液中，其中农杆菌细胞包含待瞬时表达的核苷酸序列; （b2）向步骤（b1）的浸渍大豆植物和/或其完整部分施加真空; 和（b3）释放真空，从而将溶液引入植物和/或其完整部分; 或者（c）将磨损的植物和/或其磨损的完整部分与包含农杆菌细胞和表面活性剂的溶液接触，其中农杆菌细胞包含要瞬时表达的核苷酸序列，由此核苷酸序列在一个或多个细胞中瞬时表达 的植物。
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6.发明申请公开(公告)号：US20120174262A1
公开(公告)日：2012-07-05
申请号：US13326786
申请日：2011-12-15
申请人： Kasimalai Azhakanandam , Pei Su
发明人： Kasimalai Azhakanandam , Pei Su
CPC分类号： C12N15/8205 , C12N15/8202 , C12N15/8207 , C12N15/8216 , C12N15/8257
摘要： This invention provides methods of transiently expressing a nucleotide sequence in one or more cells of a soybean plant, comprising: a) abrading the surface of a soybean plant and/or intact part thereof; and either (b1) immersing the abraded plant and/or abraded intact part thereof in a solution comprising Agrobacterium cells and a surfactant, wherein the Agrobacterium cells comprise the nucleotide sequence to be transiently expressed; (b2) applying a vacuum to the immersed plant and/or intact part thereof of step (b1); and (b3) releasing the vacuum, thereby introducing the solution into the plant and/or intact part thereof; or (c) contacting the abraded plant and/or abraded intact part thereof with a solution comprising Agrobacterium cells and a surfactant, wherein the Agrobacterium cells comprise the nucleotide sequence to be transiently expressed, whereby the nucleotide sequence is transiently expressed in one or more cells of the plant.
摘要翻译： 本发明提供了在大豆植物的一个或多个细胞中瞬时表达核苷酸序列的方法，包括：a）研磨大豆植物和/或其完整部分的表面; （b1）将磨损的植物和/或磨损的整体部分浸入包含农杆菌细胞和表面活性剂的溶液中，其中农杆菌细胞包含要瞬时表达的核苷酸序列; （b2）向步骤（b1）的浸没设备和/或其完整部分施加真空; 和（b3）释放真空，从而将溶液引入植物和/或其完整部分; 或者（c）将磨损的植物和/或其磨损的完整部分与包含农杆菌细胞和表面活性剂的溶液接触，其中农杆菌细胞包含要瞬时表达的核苷酸序列，由此核苷酸序列在一个或多个细胞中瞬时表达 的植物。
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