首页> 外国专利> Biotinated firefly luciferase, a gene for biotinated firefly luciferase, a recombinant DNA, a process for producing biotinated luciferase and a bioluminescent analysis method

Biotinated firefly luciferase, a gene for biotinated firefly luciferase, a recombinant DNA, a process for producing biotinated luciferase and a bioluminescent analysis method

机译:生物素萤火虫荧光素酶,生物素萤火虫荧光素酶的基因,重组DNA,生物素化萤光素酶的生产方法和生物发光分析方法

摘要

The present invention relates to biotinated firefly luciferase comprising a biotinated peptide and firefly luciferase linked therein, biotinated firefly luciferase having a specific amino acid sequence, a biotinated firefly luciferase gene comprising a gene coding for a biotinated peptide and a firefly luciferase gene linked therein, a biotinated firefly luciferase gene comprising a biotinated peptide gene coding for a specific amino acid sequence and a firefly luciferase gene linked therein, a recombinant DNA comprising said biotinated firefly luciferase gene inserted into a vector DNA, a process for producing biotinated firefly luciferase comprising culturing a microorganism belonging to the genus Escherichia carrying said recombinant DNA and then recovering the resulting biotinated firefly luciferase from the culture, a bioluminescent analysis method comprising using said biotinated firefly luciferase, and a bioluminescent analysis method comprising quantifying a ligand by measuring a biotinated receptor by the use of said biotinated firefly luciferase.PPAccording to the present invention, there are provided biotinated firefly luciferase, a biotinated firefly luciferase gene, a recombinant DNA, a process for producing biotinated firefly luciferase and a bioluminescent analysis method. The present invention enables the efficient production of biotinated firefly luciferase of uniform properties with the uniform structure of firefly luciferase in which one biotin molecule has been bound to a specific residue and whose activity is hardly lost by biotination and not lost even binding to streptoavidin or avidin, and the biotinated firefly luciferase of constant properties obtained in the present invention permits highly sensitive measurements in bioluminescent analysis as compared with the conventional chemically modified biotinated firefly luciferase so that the present invention is industrially extremely useful.
机译:本发明涉及一种生物素化萤火虫荧光素酶,其包含连接有生物素化肽和萤火虫荧光素酶的生物素化萤火虫荧光素酶,具有特定氨基酸序列的生物素化萤火虫荧光素酶基因,其包含编码生物素化肽的基因和萤火虫荧光素酶基因,生物素化萤火虫荧光素酶基因,其包含编码特定氨基酸序列的生物素化肽基因和与之连接的萤火虫荧光素酶基因,包含插入到载体DNA中的所述生物素化萤火虫荧光素酶基因的重组DNA,生产生物素化萤火虫荧光素酶的方法,包括培养微生物属于携带所述重组DNA然后从培养物中回收得到的生物素化萤火虫荧光素酶的埃希氏菌属的生物发光分析方法,包括使用所述生物素化萤火虫荧光素酶的生物发光分析方法,和包括通过测量定量配体的生物发光分析方法。通过使用所述生物素化萤火虫荧光素酶的生物素化受体。根据本发明,提供了生物素化萤火虫荧光素酶,生物素化萤火虫荧光素酶基因,重组DNA,生物素化萤火虫荧光素酶的生产方法和生物发光分析方法。本发明能够有效地生产具有萤火虫荧光素酶均匀结构的,性质均匀的生物素化萤火虫萤光素荧光素酶,其中一个生物素分子已经结合到特定的残基上,其活性几乎不会因生物素化作用而丧失,甚至与链霉亲和素或抗生物素蛋白结合也不会丧失。与常规的化学修饰的生物素化萤火虫萤光素酶相比,本发明获得的具有恒定特性的生物素化萤火虫萤光素酶可以在生物发光分析中进行高度灵敏的测量,因此本发明在工业上非常有用。

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