公开(公告)号：KR100733712B1

公开(公告)日：2007-06-29

申请号：KR1020060015605

申请日：2006-02-17

Applicant: 충남대학교산학협력단

Inventor： 김동명 ,  김태완 ,  박창길 ,  최차용

IPC: C12N15/52 ,  C07K1/00

Abstract: A preparation method of a cell extract useful as catalyst for synthesis of a cell-free protein and the cell-free protein synthesis method by using the same extract are provided to improve economical efficiency and productivity of the cell-free protein synthesis by simplifying the preparation process of the cell extract through high-speed centrifugation. The preparation method of the cell extract useful as catalyst for synthesis of the cell-free protein comprises the steps of: destroying the cells cultured in the medium to prepare destroyed cell solution containing cellular organelles and factors required for synthesis of a desired protein; centrifuging the destroyed cell solution at 12,000 to 30,000Xg and collecting the supernatant to prepare the cell extract; and adding the cell extract into a reaction medium containing amino acid mixture, protein synthesis energy source, gene information source and buffer solution, wherein the cell is Escherichia coli, Bacillus subtilis, wheat germ, rice germ, barley germ, CHO(Chinese hamster ovary) cell, hybridoma cell or reticulocyte; the protein synthesis energy source is ATP(adenine triphosphate), CTP(cytidine triphosphate), GTP(guanosine triphosphate), TTP(thymine triphosphate) or UTP(uridine triphosphate); and the gene information source is DNA or mRNA(messenger RNA) encoding the target protein.

Abstract translation: 提供一种可用作合成无细胞蛋白质的催化剂的细胞提取物的制备方法和使用相同提取物的无细胞蛋白质合成方法，以通过简化制备来提高无细胞蛋白质合成的经济效率和生产率 通过高速离心进行细胞提取物的过程。 用作合成无细胞蛋白质的催化剂的细胞提取物的制备方法包括以下步骤：破坏在培养基中培养的细胞以制备含有细胞器的破坏细胞溶液和合成期望蛋白质所需的因子; 以12,000至30,000Xg离心破坏的细胞溶液并收集上清液以制备细胞提取物; 将细胞提取物加入到含有氨基酸混合物，蛋白质合成能源，基因信息来源和缓冲溶液的反应介质中，其中细胞为大肠杆菌，枯草芽孢杆菌，小麦胚芽，米胚芽，大麦胚芽，CHO（中国仓鼠卵巢 ）细胞，杂交瘤细胞或网织红细胞; 蛋白质合成能源是ATP（三磷酸腺嘌呤），CTP（三磷酸胞苷），GTP（鸟苷三磷酸），TTP（三磷酸胸腺嘧啶）或UTP（尿苷三磷酸）; 而基因信息来源是编码靶蛋白的DNA或mRNA（信使RNA）。