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    • 2. 发明申请
    • Method for modifying a property of a protein
    • 改变蛋白质性质的方法
    • US20050233308A1
    • 2005-10-20
    • US10933280
    • 2004-09-03
    • Yosuke NishioEiichiro KimuraYoshihiro UsudaKazuho IkeoYoji NakamuraTakashi GojoboriYutaka KawarabayashiYumi HinoEiichi HoriJun Yamazaki
    • Yosuke NishioEiichiro KimuraYoshihiro UsudaKazuho IkeoYoji NakamuraTakashi GojoboriYutaka KawarabayashiYumi HinoEiichi HoriJun Yamazaki
    • C12N15/09C12N1/21C12N9/00C12N15/10C12P21/02C12Q1/00G01N33/48G01N33/50G06F19/00
    • C12N9/88C12N9/00C12N9/0016C12N9/1205C12N15/1034C12N15/1089
    • 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.
    • 通过以下步骤修饰蛋白质的性质:(a)从第一微生物的基因组中选择1000个或更多个基因,并从第二微生物的基因组中选择1000个或更多个基因,其中来自第一个微生物的基因是 与第二微生物的基因直系同源,其中第二微生物与第一微生物密切相关,但与第一微生物相比,在至少一个最佳生长条件下生长不同,(b)比较由第一微生物编码的氨基酸序列 基因从第一微生物到由第二微生物的直系同源基因编码的氨基酸序列,(c)检测由第一微生物的基因编码的氨基酸序列与由第二微生物的基因编码的氨基酸序列之间的取代 每对直系同源基因的微生物,(d)编码每个氨基酸取代类型的检测到的氨基酸取代,(e)cal 规定每个氨基酸取代型的频率,其中对于每个检测到的氨基酸取代类型,通过从相同的取代类型的总数中减去从第一微生物发生到第二微生物的替代类型的总数来进行校正 (f)鉴定和标记高频发生的氨基酸取代作为参与所述最佳生长条件的氨基酸取代,和(g) 将(f)中鉴定的一个或多个氨基酸取代引入编码蛋白质的基因中以改变蛋白质的性质。