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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

US4590163A Broad host range DNA cloning system for gram-negative bacteria 失效
标题翻译：用于革兰氏阴性菌的广泛的宿主范围DNA克隆系统
公开(公告)号：US4590163A
公开(公告)日：1986-05-20
申请号：US568943
申请日：1984-01-09
申请人： Donald R. Helinski , Gary S. Ditta
发明人： Donald R. Helinski , Gary S. Ditta
IPC分类号： C12N15/74 , C12N1/00 , C12N1/20 , C12N15/00 , C12P19/34 , C12P21/00 , C12P21/02 , C12P21/04
CPC分类号： C12N15/743
摘要： Novel compositions and methods for cloning DNA in a broad range of Gram-negative bacteria. Relatively small vectors are prepared substantially free of DNA sequences coding for transfer functions, while having a broad host range replication function. While the plasmid is not self-transmissible, it is transmissible by means of a helper plasmid, which desirably has a narrow host range replication function. Exogenous DNA may be inserted into the first plasmid and conjugally transferred to the target host by means of a convenient bacterial host.
摘要翻译：用于在广泛的革兰氏阴性细菌中克隆DNA的新型组合物和方法。制备基本上不含编码转移功能的DNA序列的相对小的载体，同时具有广泛的宿主范围复制功能。虽然质粒不是自传播的，但是其可以通过辅助质粒传递，其需要具有窄的宿主范围复制功能。可以将外源DNA插入到第一个质粒中，并通过方便的细菌宿主将其偶联转移到靶宿主体内。

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