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1. 发明授权

US5583024A Recombinant expression of Coleoptera luciferase 失效
标题翻译：鞘翅目萤光素酶的重组表达
公开(公告)号：US5583024A
公开(公告)日：1996-12-10
申请号：US60091
申请日：1993-05-10
申请人： Marlene D. McElroy, deceased , W. D. McElroy, executor , Donald R. Helinski , Keith V. Wood , Jeffrey R. De Wet , David W. Ow , Stephen H. Howell
发明人： Marlene D. McElroy, deceased , W. D. McElroy, executor , Donald R. Helinski , Keith V. Wood , Jeffrey R. De Wet , David W. Ow , Stephen H. Howell
IPC分类号： C12N1/21 , C12N9/02 , C12N9/38 , C12N15/52 , C12Q1/68 , G01N33/535 , G01N33/573 , C12N15/53 , C12N15/63
CPC分类号： C12N9/0069 , C12N15/52 , C12Q1/6897 , C12Y113/12007 , C12Y302/01023 , G01N33/535 , G01N33/573 , C07K2319/00 , C07K2319/61
摘要： A method is disclosed for producing a protein which expresses bioluminescence activity which involves combining two polydeoxyribonucleotides, one containing a continuous sequence of codons encoding a polypeptide which comprises a single covalently bonded molecular structure and which catalyzes the oxidation of insect luciferin to yield light and the other which causes DNA transcription, and obtaining the polypeptide by transcription and subsequent translation. The insect luciferin is derived from bioluminescent insect, preferably Diptera and Coleoptera (fireflies and beetles). Hybrid proteins are similarly formed by inclusion of an additional polydeoxyribonucleotide encoding for a second polypeptide such that their respective polypeptide-encoding reading frames form a continuous reading frame. Also disclosed is a method for quantitatively assaying a fluid for the presence of an unknown quantity of antigen using the hybrid proteins bonded to antibody directed against the antigen, determining luminescence after reaction of the antibody and antigen, and determining the amount of the antigens by comparison of the determined luminescence with the luminescence previously measured under equivalent conditions for equivalent hybrid proteins reacted under equivalent conditions with known amounts of the antigen.
摘要翻译：公开了用于产生表达生物发光活性的蛋白质的方法，其涉及组合两种多脱氧核糖核苷酸，一种含有编码多肽的密码子的连续序列，其包含单个共价键合的分子结构并且催化昆虫萤光素的氧化以产生光，另一种其导致DNA转录，并通过转录和随后的翻译获得多肽。昆虫萤光素衍生自生物发光昆虫，优选双翅目和鞘翅目（萤火虫和甲虫）。类似地，通过包含编码第二多肽的另外的多脱氧核糖核苷酸使得它们各自的多肽编码阅读框形成连续的阅读框，类似地形成杂交蛋白。还公开了使用与针对抗原的抗体结合的杂交蛋白定量测定流体存在未知量的抗原的方法，测定抗体和抗原反应后的发光，并通过比较确定抗原的量的等效杂交蛋白在等同条件下在已知量的抗原的等同条件下反应的在先前测量的发光的确定的发光。

2. 发明授权

US5700673A Recombinantly produced Coleoptera luciferase and fusion proteins thereof 失效
标题翻译：重组生产的鞘翅目荧光素酶及其融合蛋白
公开(公告)号：US5700673A
公开(公告)日：1997-12-23
申请号：US458828
申请日：1995-06-02
申请人： Marlene D. McElroy, deceased , Donald R. Helinski , Keith V. Wood , Jeffrey R. De Wet , David W. Ow , Stephen H. Howell
发明人： Marlene D. McElroy, deceased , Donald R. Helinski , Keith V. Wood , Jeffrey R. De Wet , David W. Ow , Stephen H. Howell
IPC分类号： C12N1/21 , C12N9/02 , C12N9/38 , C12N15/52 , C12Q1/68 , G01N33/535 , G01N33/573
CPC分类号： C12N9/0069 , C12N15/52 , C12Q1/6897 , C12Y113/12007 , C12Y302/01023 , G01N33/535 , G01N33/573 , C07K2319/00 , C07K2319/61
摘要： A method is disclosed for producing a protein which expresses bioluminescence activity which involves combining two polydeoxyribonucleotides, one containing a continuous sequence of codons encoding a polypeptide which comprises a single covalently bonded molecular structure and which catalyzes the oxidation of insect luciferin to yield light and the other which causes DNA transcription, and obtaining the polypeptide by transcription and subsequent translation. The insect luciferin is derived from bioluminescent insect, preferably Diptera and Coleoptera (fireflies and beetles). Hybrid proteins are similarly formed by inclusion of an additional polydeoxyribonucleotide encoding for a second polypeptide such that their respective polypeptide-encoding reading frames form a continuous reading frame. Also disclosed is a method for quantitatively assaying a fluid for the presence of an unknown quantity of antigen using the hybrid proteins bonded to antibody directed against the antigen, determining luminescence after reaction of the antibody and antigen, and determining the amount of the antigens by comparison of the determined luminescence with the luminescence previously measured under equivalent conditions for equivalent hybrid proteins reacted under equivalent conditions with known amounts of the antigen.
摘要翻译：公开了用于产生表达生物发光活性的蛋白质的方法，其涉及组合两种多脱氧核糖核苷酸，一种含有编码多肽的密码子的连续序列，其包含单个共价键合的分子结构并且催化昆虫萤光素的氧化以产生光，另一种其导致DNA转录，并通过转录和随后的翻译获得多肽。昆虫萤光素衍生自生物发光昆虫，优选双翅目和鞘翅目（萤火虫和甲虫）。类似地，通过包含编码第二多肽的另外的多脱氧核糖核苷酸使得它们各自的多肽编码阅读框形成连续的阅读框，类似地形成杂交蛋白。还公开了使用与针对抗原的抗体结合的杂交蛋白定量测定流体存在未知量的抗原的方法，测定抗体和抗原反应后的发光，并通过比较确定抗原的量的等效杂交蛋白在等同条件下在已知量的抗原的等同条件下反应的在先前测量的发光的确定的发光。

3. 发明授权

US4407956A Cloned cauliflower mosaic virus DNA as a plant vehicle 失效
标题翻译：克隆花椰菜花叶病毒DNA作为植物载体
公开(公告)号：US4407956A
公开(公告)日：1983-10-04
申请号：US243330
申请日：1981-03-13
申请人： Stephen H. Howell
发明人： Stephen H. Howell
IPC分类号： C12N15/82 , C12N15/00 , C12N1/00 , C12N5/00
CPC分类号： C12N15/8203 , Y10S47/01
摘要： Novel methods and compositions are provided for preparing vectors for the introduction of DNA into plant cells for transcription and expression of the DNA. Particularly, cauliflower mosaic virus DNA is inserted into a bacterial cloning vehicle to provide a recombinant plasmid for cloning in a microorganism. The resulting cloned plasmid is genetically manipulated to introduce exogenous or heterologous DNA. Conveniently, linkers can be inserted which provide for a unique restriction site for insertion of exogenous or heterologous DNA. At each stage the modified plasmid may be cloned to provide for relatively large amounts of material for modification and isolation. Besides insertions, deletions may be made, removing non-essential portions of the virus. After completion of the viral modifications, the CaMV is excised from the hybrid plasmid by restriction enzyme cleavage and may be used for systemic infection of plants.
摘要翻译：提供了新的方法和组合物，用于制备用于将DNA引入植物细胞以用于转录和表达DNA的载体。特别地，将花椰菜花叶病毒DNA插入细菌克隆载体中以提供用于在微生物中克隆的重组质粒。遗传操作得到的克隆质粒引入外源或异源DNA。方便地，可以插入接头，其提供用于插入外源或异源DNA的唯一限制性位点。在每个阶段，可以克隆修饰的质粒以提供相对大量的用于修饰和分离的材料。除了插入之外，可以进行删除，去除病毒的非必需部分。病毒修饰完成后，通过限制酶切割从杂交质粒切除CaMV，可用于植物的全身感染。

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