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公开(公告)号：US07632936B2
公开(公告)日：2009-12-15
申请号：US11459527
申请日：2006-07-24
CPC分类号： C12N9/22 , C12N15/8218 , C12N15/8283
摘要： The present invention includes a method for detecting and isolating sugarcane proteins that interact with the HC-Pro and P1 proteins of SrMV and other proteins involved in gene silencing, particularly in sugarcane. The method uses a two hybrid assay with an HC-Pro, P1, or other silencing-related protein-containing bait protein and a prey protein containing a polypeptide encoded by a DNA molecule in a cDNA library. The method also includes identification of false positives through reverse two-hybrid assays and using in vitro techniques such as farwestern blots or pull down assays where plant physiological conditions may be replicated. Finally, interactions may be confirmed in planta. Some novel proteins used in and discovered using the these methods are also identified. Methods of using viral and plant proteins to regulate silencing in plants such as sugarcane are also discussed.
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2.发明授权Isolation of proteins involved in posttranscriptional gene silencing and methods of use 有权分离参与转录后基因沉默的蛋白质和使用方法公开(公告)号：US07534614B2
公开(公告)日：2009-05-19
申请号：US11460815
申请日：2006-07-28
CPC分类号： C12N9/22 , C12N15/8218 , C12N15/8283
摘要： The present invention discloses transgenic plant cells transformed with a nucleic acid encoding a SrMV P1/HC-Pro protein, wherein expression of the protein suppresses post-transcriptional gene silencing in the cell. The transgenic plant cell may be located in an embryonic callus or in a mature plant. The plant may be a monocot plant.
摘要翻译： 本发明公开了用编码SrMV P1 / HC-Pro蛋白的核酸转化的转基因植物细胞，其中蛋白质的表达抑制细胞中的转录后基因沉默。 转基因植物细胞可以位于胚胎愈伤组织或成熟植物中。 植物可以是单子叶植物。
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3.发明申请ISOLATION OF PROTEINS INVOLVED IN POSTTRANSCRIPTIONAL GENE SILENCING AND METHODS OF USE 审中-公开分离基因沉默中涉及的蛋白质的分离及其使用方法公开(公告)号：US20120278946A1
公开(公告)日：2012-11-01
申请号：US12817309
申请日：2010-06-17
IPC分类号： C12N15/82 , C12N9/16 , C12N5/10 , C07K14/08 , C07K14/415
CPC分类号： C12N9/22 , C12N15/8218 , C12N15/8283
摘要： The present disclosure relates, according to some embodiments, to methods of increasing viral resistance in a plant, methods of increasing transgene expression in a plant, and methods of suppressing expression of a native gene in a plant. In some embodiments, the disclosure also relates to polypeptides involved in post-translational gene silencing (PTGS), for e.g., a protein active in PTGS or a suppressor of PTGS. The disclosure further relates to transgenic plant cells in which PTGS is altered (e.g., enhanced or suppressed) according to some embodiments.
摘要翻译： 根据一些实施方案，本公开涉及增加植物中病毒抗性的方法，增加植物中转基因表达的方法，以及抑制植物中天然基因表达的方法。 在一些实施方案中，本公开还涉及涉及翻译后基因沉默（PTGS）的多肽，例如在PTGS中活化的蛋白质或PTGS抑制剂。 本公开还涉及根据一些实施方案其中PTGS被改变（例如增强或抑制）的转基因植物细胞。
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4.发明授权Isolation of proteins involved in posttranscriptional gene silencing and methods of use 有权分离参与转录后基因沉默的蛋白质和使用方法公开(公告)号：US07777096B2
公开(公告)日：2010-08-17
申请号：US11460836
申请日：2006-07-28
IPC分类号： C12N15/82
CPC分类号： C12N9/22 , C12N15/8218 , C12N15/8283
摘要： The present disclosure includes methods of using viral and plant proteins to regulate silencing in plants such as monocots. Some embodiments include methods of increasing expression of transgenes in plants. Such methods may include selecting a protein involved in post-translational gene silencing (PTGS), for e.g., a protein active in PTGS or a suppressor of PTGS, in a plant and transforming the plant with a nucleic acid encoding the protein involved in PTGS wherein expression of the protein involved in PTGS increases expression of the transgene in the plant. One non-limiting example of such a protein is the Sorghum Mosaic Virus (SrMV) P1/HC-Pro. Methods of the disclosure may be used to regulate silencing of transgenes and to increase expression of transgenes in a variety of plants such as sugarcane, corn, sorghum, and/or rice.
摘要翻译： 本公开内容包括使用病毒和植物蛋白调节植物如单子叶植物中的沉默的方法。 一些实施方案包括增加植物中转基因表达的方法。 这样的方法可以包括选择参与翻译后基因沉默（PTGS）的蛋白质，例如在植物中PTGS或PTGS抑制剂中的活性蛋白质，以及用编码PTGS参与的蛋白质的核酸转化植物，其中 参与PTGS的蛋白质的表达增加植物中转基因的表达。 这种蛋白质的一个非限制性实例是高粱花叶病毒（SrMV）P1 / HC-Pro。 本公开的方法可用于调节转基因的沉默并增加多种植物如甘蔗，玉米，高粱和/或水稻中转基因的表达。
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5.发明授权Isolation of proteins involved in posttranscriptional gene silencing and methods of use 有权分离参与转录后基因沉默的蛋白质和使用方法公开(公告)号：US07632937B2
公开(公告)日：2009-12-15
申请号：US11459535
申请日：2006-07-24
CPC分类号： C12N9/22 , C12N15/8218 , C12N15/8283
摘要： The present invention includes a method for detecting and isolating sugarcane proteins that interact with the HC-Pro and P1 proteins of SrMV and other proteins involved in gene silencing, particularly in sugarcane. The method uses a two hybrid assay with an HC-Pro, P1, or other silencing-related protein-containing bait protein and a prey protein containing a polypeptide encoded by a DNA molecule in a cDNA library. The method also includes identification of false positives through reverse two-hybrid assays and using in vitro techniques such as farwestern blots or pull down assays where plant physiological conditions may be replicated. Finally, interactions may be confirmed in planta. Some novel proteins used in and discovered using the these methods are also identified. Methods of using viral and plant proteins to regulate silencing in plants such as sugarcane are also discussed.
摘要翻译： 本发明包括用于检测和分离与参与基因沉默的SrMV和其他蛋白质的HC-Pro和P1蛋白质相互作用的甘蔗蛋白质的方法，特别是在甘蔗中。 该方法使用具有HC-Pro，P1或其它沉默相关蛋白的诱饵蛋白质的两种杂交测定法和含有cDNA文库中的DNA分子编码的多肽的猎物蛋白质。 该方法还包括通过反向双杂交测定法鉴定假阳性并使用体外技术，例如可以复制植物生理条件的远紫外线印迹或下拉测定法。 最后，在植物中可以确认相互作用。 还鉴定了使用这些方法使用和发现的一些新型蛋白质。 还讨论了使用病毒和植物蛋白调节植物如甘蔗中的沉默的方法。
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公开(公告)号：US07001739B2
公开(公告)日：2006-02-21
申请号：US10226715
申请日：2002-08-23
IPC分类号： G01N33/569 , G01N33/53 , C12N15/33
CPC分类号： C12N9/22 , C12N15/8218 , C12N15/8283
摘要： The present invention includes a method for detecting and isolating sugarcane proteins that interact with the HC-Pro and P1 proteins of SrMV and other proteins involved in gene silencing, particularly in sugarcane. The method uses a two hybrid assay with an HC-Pro, P1, or other silencing-related protein-containing bait protein and a prey protein containing a polypeptide encoded by a DNA molecule in a cDNA library. The method also includes identification of false positives through reverse two-hybrid assays and using in vitro techniques such as farwestern blots or pull down assays where plant physiological conditions may be replicated. Finally, interactions may be confirmed in planta. Some novel proteins used in and discovered using the these methods are also identified. Methods of using viral and plant proteins to regulate silencing in plants such as sugarcane are also discussed.
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