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1. 发明申请

WO2006003018A2 BIOLOGICALLY SAFE TRANSIENT PROTEIN EXPRESSION IN PLANTS 审中-公开
标题翻译：生物安全瞬态蛋白表达在植物中
公开(公告)号：WO2006003018A2
公开(公告)日：2006-01-12
申请号：PCT/EP2005/007361
申请日：2005-07-07
申请人： ICON GENETICS AG , MARILLONNET, Sylvestre , ENGLER, Carola , MÜHLBAUER, Stefan , HERZ, Stefan , WERNER, Stefan , KLIMYUK, Victor , GLEBA, Yuri
发明人： MARILLONNET, Sylvestre , ENGLER, Carola , MÜHLBAUER, Stefan , HERZ, Stefan , WERNER, Stefan , KLIMYUK, Victor , GLEBA, Yuri
IPC分类号： C12N15/82 , A01H5/12 , C12N15/74
CPC分类号： C12N15/8203 , C12N15/8205 , C12N15/8216 , C12N15/8257
摘要： A process of producing a protein of interest by expression of said protein of interest from a sequence of interest in a plant or in plant leaves, comprising: a) transfecting said plant or said plant leaves by infiltrating said plant or said plant leaves with an Agrobacterium strain in the presence of a complementing factor, said Agrobacterium strain containing in T-DNA a heterelogous DNA sequence having a sequence portion encoding a replicon, wherein said sequence encoding a replicon contains sequences necessary for replicon function of said replicon, said sequences being derived from a plant virus, and said sequence of interest to be expressed from said replicon, b) optionally isolating said protein of interest from said plant or said plant leaves infiltrated in step (a), wherein said Agrobacterium strain is provided with a first genetic modification rendering said Agrobacterium strain defective for transfecting organisms with said T-DNA in the absence of said complementing factor.
摘要翻译：通过从植物或植物叶中感兴趣的序列表达所述感兴趣的蛋白质来产生感兴趣的蛋白质的方法，包括：a）通过用农杆菌浸润所述植物或所述植物叶来转染所述植物或所述植物叶子所述含有T-DNA的所述农杆菌菌株具有编码复制子的序列部分的恒耳DNA序列，其中所述编码复制子的序列含有所述复制子的复制子功能所需的序列，所述序列衍生自所述复制子表达的所述感兴趣的序列，b）任选地从所述植物或所述在步骤（a）中渗透的所述植物叶分离所述目的蛋白质，其中所述农杆菌菌株被提供第一遗传修饰呈现所述农杆菌菌株在没有所述补体因子的情况下缺陷用于用所述T-DNA转染生物体。

2. 发明申请

WO2009109360A1 METHOD OF PROTEASE PRODUCTION IN PLANTS 审中-公开
标题翻译：在植物中生产蛋白质的方法
公开(公告)号：WO2009109360A1
公开(公告)日：2009-09-11
申请号：PCT/EP2009/001502
申请日：2009-03-03
申请人： ICON GENETICS GMBH , KANDZIA, Romy , ENGLER, Carola , MARILLONNET, Sylvestre , KLIMYUK, Victor , GLEBA, Yuri
发明人： KANDZIA, Romy , ENGLER, Carola , MARILLONNET, Sylvestre , KLIMYUK, Victor , GLEBA, Yuri
IPC分类号： C12N15/82 , A01H5/00
CPC分类号： C12N15/8257 , C07K14/415
摘要： A process of producing a protease in a plant or in plant cells, comprising (a) providing a plant comprising a heterologous nucleotide sequence comprising a coding sequence encoding a fusion protein, said fusion protein comprising: an apoplast or plastid signal peptide; a SUMO protein or a derivative of a SUMO protein; and a zymogen of said protease, and (b) expressing said fusion protein.
摘要翻译：一种在植物或植物细胞中产生蛋白酶的方法，包括（a）提供包含异源核苷酸序列的植物，所述异源核苷酸序列包含编码融合蛋白的编码序列，所述融合蛋白包含：质外体或质体信号肽; SUMO蛋白或SUMO蛋白的衍生物; 和所述蛋白酶的酶原，和（b）表达所述融合蛋白。

3. 发明申请

WO2008028661A2 POTEXVIRUS- DERIVED REPLICON 审中-公开
标题翻译： POTEXVIRUS-衍生的REPLICON
公开(公告)号：WO2008028661A2
公开(公告)日：2008-03-13
申请号：PCT/EP2007/007785
申请日：2007-09-06
申请人： ICON GENETICS GMBH , MARILLONNET, Sylvestre , ENGLER, Carola , KLIMYUK, Victor , GLEBA, Yuri
发明人： MARILLONNET, Sylvestre , ENGLER, Carola , KLIMYUK, Victor , GLEBA, Yuri
IPC分类号： C12N15/82
CPC分类号： C12N15/8203 , C12N15/8216 , C12N2770/26011 , C12N2770/26043
摘要： Nucleic acid comprising or encoding an RNA replica comprising, in this order, the following segments (i) to (iii): i) a nucleic acid sequence encoding a potexvirus RNA-dependent RNA polymerase or a function-conservative variant thereof; ii) a nucleic acid sequence comprising: a) a potexvirus triple gene block or a function-conservative variant thereof and b) a sequence encoding a potexviral coat protein or a function-conservative variant thereof; or a sequence encoding a tobago viral movement protein; and iii) a heterologous nucleic acid sequence expressible from said replica in a plant or in plant tissue.
摘要翻译：包含或编码RNA复制品的核酸，其依次包括以下片段（i）至（iii）：i）编码病毒RNA依赖性RNA聚合酶或其功能保守变体的核酸序列; ii）核酸序列，其包含：a）罐状病毒三重基因嵌段或其功能保守变体，和b）编码潜在外壳蛋白或其功能保守变体的序列; 或编码多巴胺病毒运动蛋白的序列; 和iii）在植物或植物组织中可从所述复制品表达的异源核酸序列。

4. 发明申请

WO2011154147A1 SYSTEM AND METHOD OF MODULAR CLONING 审中-公开
标题翻译：模块化克隆的系统和方法
公开(公告)号：WO2011154147A1
公开(公告)日：2011-12-15
申请号：PCT/EP2011/002843
申请日：2011-06-09
申请人： ICON GENETICS GMBH , WEBER, Ernst , WERNER, Stefan , ENGLER, Carola , GRUETZNER, Ramona , MARILLONNET, Sylvestre
发明人： WEBER, Ernst , WERNER, Stefan , ENGLER, Carola , GRUETZNER, Ramona , MARILLONNET, Sylvestre
IPC分类号： C12N15/66
CPC分类号： C12N15/66 , C12N15/1093
摘要： System for producing a nucleic acid construct of interest, said system comprising: a set of n entry DNAs numbered 1 to n, n being an integer of at least 2, each of said n entry DNAs comprising in this order: (i) a type lIs restriction endonuclease recognition site followed by the cleavage site thereof; (ii) a sequence portion linking the cleavage site of said recognition site of item (i) with the cleavage site of the recognition site of the following item (iii), and (iii) a cleavage site of a further type Ms restriction endonuclease recognition site followed by the recognition site of said cleavage site; the cleavage sites of the type lIs restriction endonuclease recognition sites of item (iii) of entry DNAs 1 to n-1 are complementary to the cleavage sites of the type lIs restriction endonuclease recognition sites of item (i) of entry DNAs 2 to n, respectively; the cleavage site of the type Ms restriction endonuclease recognition site of item (iii) of entry DNA n is complementary to the cleavage site of the type lIs restriction endonuclease recognition site of item (i) of entry DNA 1 for allowing annealing of complementary single-stranded overhangs formed by restriction at recognition site (i) of entry DNA 1 and at recognition site (iii) of entry DNA n; said system further comprising a destination vector comprising in this order: (I) a type Ms restriction endonuclease recognition site followed by the cleavage site thereof; (II) a vector backbone preferably comprising a selectable marker gene, said vector backbone linking the cleavage sites of said recognition sites of items (I) and the following item (III); (III) a further cleavage site of a type Ms restriction endonuclease recognition site followed by the recognition site of said cleavage site, and (IV) optionally, an insert between the recognition sites of item (III) and item (i); said cleavage sites of items (I) and (III) being different and non-complementary, said recognition sites of items (I) and (III) being preferably recognitions sites of the same endonuclease.
摘要翻译：用于产生感兴趣的核酸构建体的系统，所述系统包括：一组n至n的n个入口DNA，n为至少2的整数，所述n个入口DNA中的每一个按以下顺序包括：（i） lIs限制性内切核酸酶识别位点，然后是其切割位点; （ii）将项目（i）的所述识别位点的切割位点与下述（iii）的识别位点的切割位点连接的序列部分，和（iii）另外类型的Ms限制性内切核酸酶识别的切割位点其后是所述切割位点的识别位点; 入口DNA1至n-1项目（ⅲ）的III型限制性内切核酸酶识别位点的切割位点与条目DNA2至n的第（i）项的第II型限制性内切核酸酶识别位点的切割位点互补，分别; 入口DNA n项目（iii）的Ms限制性内切核酸酶识别位点的切割位点与用于允许互补单链DNA退火的入口DNA 1的第（i）项的I型限制性内切核酸酶识别位点的切割位点互补，在入口DNA 1的识别位点（i）和进入DNA n的识别位点（iii）处形成的疏水突出物; 所述系统还包括目的载体，其包含以下顺序：（I）Ms型限制性内切核酸酶识别位点，其后是其切割位点; （II）优选包含选择性标记基因的载体骨架，所述载体骨架连接项目（I）的所述识别位点的切割位点和下列项目（III）; （III）另外的Ms限制性内切核酸酶识别位点的切割位点，随后是所述切割位点的识别位点，和（IV）任选地，（III）和（i）项的识别位点之间的插入片段; 所述项目（I）和（III）的切割位点是不同的和不相互补充的，所述项目（I）和（III）的所述识别位点优选为相同核酸内切酶的识别位点。

5. 发明申请

WO2010040531A2 PROCESS OF CLEAN CLONING 审中-公开
标题翻译：清洁克隆过程
公开(公告)号：WO2010040531A2
公开(公告)日：2010-04-15
申请号：PCT/EP2009/007237
申请日：2009-10-08
申请人： ICON GENETICS GMBH , MARILLONNET, Sylvestre , WERNER, Stefan , ENGLER, Carola , KANDZIA, Romy , THIEME, Frank , WEBER, Ernst
发明人： MARILLONNET, Sylvestre , WERNER, Stefan , ENGLER, Carola , KANDZIA, Romy , THIEME, Frank , WEBER, Ernst
IPC分类号： C12N15/09 , C12N15/10
CPC分类号： C12N15/64 , C12N15/66
摘要： A process of inserting a nucleic acid sequence of interest into an acceptor nucleic acid, comprising the following steps: amplifying by PCR a DNA comprising in the following order a sequence segment U, a nucleic acid sequence segment of known nucleotide sequence K2 and a nucleic acid sequence segment of known sequence K3 using a forward primer defining a first end of the amplified DNA and a reverse primer defining a second end of the amplified DNA, said reverse primer terminating at its 3'-end in a nucleotide sequence of nucleic acid sequence segment K3; treating the linear double-stranded DNA molecules contained in the PCR product obtained in the previous step with an exonuclease to obtain a single-stranded overhang at the first end of the DNA and a single-stranded overhang comprising nucleic acid segments K2 and K3 at the second end of the DNA; and annealing the product of the previous step to a linearized double-stranded acceptor nucleic acid having at a first end thereof a single-stranded overhang complementary to the single-stranded overhang of the first end of the DNA and at a second end thereof a single- stranded overhang complementary to the single-stranded sequence segment K2 of the second end of the DNA.
摘要翻译：将感兴趣的核酸序列插入受体核酸中的方法，包括以下步骤：通过PCR扩增包含以下顺序的DNA的序列：序列区段U，核酸序列片段使用限定扩增的DNA的第一末端的正向引物和限定扩增的DNA的第二末端的反向引物的已知核苷酸序列K2和已知序列K3的核酸序列区段，所述反向引物在其3'末端终止于核酸序列片段K3的核苷酸序列; 用外切核酸酶处理在前一步骤中获得的PCR产物中所含的线性双链DNA分子，以在DNA的第一末端获得单链突出端和单链突出端，所述单链突出端包含位于第一端的核酸片段K2和K3 DNA的第二末端; 和将前一步骤的产物退火成线性化双链受体核酸，所述线性化双链受体核酸在其第一末端具有与DNA第一末端的单链突出端互补的单链突出端，并且在其第二末端具有单一突出端 - 与DNA第二末端的单链序列片段K2互补的多余突出端。

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