申请人:Nishio Yosuke
公开号:US20050233308A1
公开(公告)日:2005-10-20
A property of a protein is modified by the following steps: (a) selecting 1000 or more genes from the genome of a first microorganism, and selecting 1000 or more genes from the genome of a second microorganism, wherein the genes from the first microorganism are orthologs to the genes from the second microorganism, and wherein the second microorganism is closely related to the first microorganism, but grows differently under at least one optimum growth condition when compared with the first microorganism, (b) comparing an amino acid sequence encoded by a gene from the first microorganism to an amino acid sequence encoded by the orthologous gene from the second microorganism, (c) detecting substitutions between the amino acid sequence encoded by a gene from the first microorganism and the amino acid sequence encoded by a gene from the second microorganism for each pair of orthologous genes, (d) compiling the detected amino acid substitutions for each amino acid substitution type, (e) calculating the frequency of each amino acid substitution type, wherein for each detected amino acid substitution type, a correction is made by subtracting the total number of substitution types which occur from the first microorganism to the second microorganism from the total number of the same substitution type which occurs in the reverse direction, or from the second microorganism to the first microorganism, (f) identifying and labelling the amino acid substitutions which occur at a high frequency as amino acid substitutions which are involved in said optimum growth condition, and (g) introducing one or more of the amino acid substitutions identified in (f) into the gene encoding the protein to modify a property of the protein.
蛋白质的一个特性通过以下步骤进行修改:(a)从第一个微生物的基因组中选择1000个或更多基因,从第二个微生物的基因组中选择1000个或更多基因,其中来自第一个微生物的基因与来自第二个微生物的基因是同源基因,在第二个微生物与第一个微生物密切相关,但在至少一个最佳生长条件下与第一个微生物生长方式不同,(b)将由第一个微生物的基因编码的氨基酸序列与由同源基因编码的第二个微生物的氨基酸序列进行比较,(c)检测每对同源基因之间编码的氨基酸序列之间的替换,(d)编译每种氨基酸替换类型的检测到的氨基酸替换,(e)计算每种氨基酸替换类型的频率,其中对于每种检测到的氨基酸替换类型,通过从第一个微生物到第二个微生物发生的替换类型总数减去在相反方向上发生的相同替换类型的总数,或从第二个微生物到第一个微生物,进行校正,(f)识别和标记高频发生的氨基酸替换作为涉及所述最佳生长条件的氨基酸替换,(g)将在(f)中确定的一个或多个氨基酸替换引入编码蛋白质的基因中,以修改蛋白质的特性。