首页> 外国专利> METHOD FOR THE IN VIVO MODIFICATION OF THE SYNTHESIS ACTIVITY OF A METABOLITE BY MEANS OF THE MODIFICATION OF A GENE THE ACTIVITY OF WHICH IS NOT THE ORIGINAL ACTIVITY

METHOD FOR THE IN VIVO MODIFICATION OF THE SYNTHESIS ACTIVITY OF A METABOLITE BY MEANS OF THE MODIFICATION OF A GENE THE ACTIVITY OF WHICH IS NOT THE ORIGINAL ACTIVITY

机译:通过基因修饰而不是原始活性活体内修饰代谢产物合成活性的方法

摘要

The invention relates to a method for altering a protein X such as to modify the characteristics thereof by a) obtaining the mutants X* of the sequence coding for protein X, by means of aleatory mutagenesis, b) transformation of cells with a phenotype [P-] with vectors comprising the mutated nucleic acids obtained in step (a) which code for proteins X*, where P- signifies that said cells are auxotrophic for substance P, P begin the product of the action of X on the natural substrate thereof S, c) culturing said cells in a medium comprising a substrate S*, S* being an analogue of the natural substrate S of the protein X, d) selection of the cells [P-:: X*] which have survived step c) in which the proteins X* can biosynthesise the product P from the substrate S*. The invention further relates to mutated proteins X, nucleic acids, expression vectors, host cells comprising a vector, use of N-dideoxyribosyl transferases for the transfer of a dideoxyribose (ddR) from a dideoxyribonucleoside to another nucleoside, a method for production of compounds comprising a step using a mutated protein and a strain of E. coli.
机译:本发明涉及一种改变蛋白质X以改变其特性的方法,该方法是:a)通过偶然诱变获得编码蛋白质X的序列的突变体X *,b)转化具有表型[P]的细胞-]含有包含在步骤(a)中获得的编码蛋白质X *的突变核酸的载体,其中P-表示所述细胞是物质P的营养缺陷型,P开始X在其天然底物S上的作用产物,c)在包含底物S *的培养基中培养所述细胞,S *是蛋白质X的天然底物S的类似物,d)选择在步骤c)中存活的细胞[P-:: X *]其中蛋白质X *可以从底物S *生物合成产物P。本发明进一步涉及突变的蛋白质X,核酸,表达载体,包含该载体的宿主细胞,N-二脱氧核糖基转移酶用于将双脱氧核糖(ddR)从双脱氧核糖核苷转移至另一核苷的用途,制备化合物的方法,包括使用突变蛋白和大肠杆菌菌株的步骤。

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