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11.发明公开목적하는 단백질 발현 수준을 제공하는 초기 코돈의발굴방법, 및 이를 이용한 재조합 단백질의 발현 조절 방법및 생산 방법 有权用于筛选提供所需表达蛋白质水平的初始编码的方法，以及用于调节表达和产生重组蛋白的方法
公开(公告)号：KR1020090097056A
公开(公告)日：2009-09-15
申请号：KR1020080022217
申请日：2008-03-10
Applicant: 충남대학교산학협력단
CPC classification number: C12N15/67 , C12N15/102 , C12N15/74
Abstract: A method for screening an initial codon, which provides target protein level is provided to express an active protein at high speed and synthesize signal peptide having expression intensity by regulating the expression intensity of signal peptide. A method for screening an initial codon, which provides expression level of target protein comprises: a step of changing the target gene in one or more site of +2 to +13 codon in random; a step of isolating individual gene and amplifying the isolated gene through colony PCR; a step of performing cell-free protein synthesis; and a step of selecting a codon by irradiating.
Abstract translation: 提供了提供靶蛋白水平的初始密码子筛选方法，以高速表达活性蛋白，通过调节信号肽的表达强度合成具有表达强度的信号肽。 用于筛选提供靶蛋白表达水平的初始密码子的方法包括：随机改变+2至+13个密码子的一个或多个位点中的靶基因的步骤; 分离单个基因并通过菌落PCR扩增分离的基因的步骤; 进行无细胞蛋白质合成的步骤; 以及通过照射选择密码子的步骤。
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公开(公告)号：KR101671688B1
公开(公告)日：2016-11-03
申请号：KR1020150083116
申请日：2015-06-12
Applicant: 충남대학교산학협력단
CPC classification number: G01N33/84 , C07K1/02 , C12P21/02 , C12Y401/01 , G01N33/6845
Abstract: 본발명은효소를이용하여 pH를조절하는것을특징으로하는무세포단백질합성방법에관한것으로, 더욱상세하게는아미노산탈카르복실화효소를이용하여 ATP의재생시 생성되는수소이온을제거함으로써, pH를조절하는것을특징으로하는무세포단백질합성방법에관한것이다. 본발명의무세포단백질합성방법은단백질의발현량을향상시킬뿐만아니라발현된단백질의분리정제없이바로활성분석에활용할수 있는이점이있다.
Abstract translation: 无细胞蛋白质合成的方法的特征在于通过使用酶来控制pH。 例如，通过使用氨基酸脱羧酶，根据ATP再生期间产生的氢离子的去除来控制pH。 本发明的无细胞蛋白质合成方法的优点在于，不仅蛋白质的表达量增加，而且表达的蛋白质也可以直接用于活性分析，而不进行任何分离或纯化。
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公开(公告)号：KR1020140098469A
公开(公告)日：2014-08-08
申请号：KR1020130011153
申请日：2013-01-31
Applicant: 충남대학교산학협력단
Abstract: The present invention relates to a gel matrix comprising a hydrogel holding microbeads, on which a first capturer conjugated with a target gene template and a second capturer conjugated with Ni-NTA (nitrilotriacetic acid) are combined, for synthesizing in situ protein, a method for manufacturing the gel matrix, protein in situ array according to the method, and a method for detecting or analyzing a protein using the same. The gel matrix according to the present invention allows one or multiple genes to synthesize proteins rapidly and also does not need complicated separation and purification processes. Additionally, the present invention enables a three-dimensional structure of the protein to be maintained, thereby being useful for detection and analysis of proteins including antigen-antibody reaction. Also, the present invention enables conjugation of various kinds of genes so that various proteins are synthesized at the same time, thereby being useful for researches and industries related to protein such as improvement of proteins, synthesis of biomolecules, molecular diagnosis, and synthetic biology.
Abstract translation: 本发明涉及一种凝胶基质，其包含保持微珠的水凝胶，其上结合有靶基因模板的第一捕获剂和与Ni-NTA（次氮基三乙酸）缀合的第二捕获剂用于合成原位蛋白， 根据该方法制造凝胶基质，原位阵列蛋白，以及使用其进行蛋白质的检测或分析的方法。 根据本发明的凝胶基质允许一个或多个基因快速合成蛋白质，也不需要复杂的分离和纯化过程。 此外，本发明能够保持蛋白质的三维结构，从而可用于检测和分析包括抗原 - 抗体反应的蛋白质。 此外，本发明能够使各种基因的缀合使得各种蛋白质同时合成，从而可用于与蛋白质相关的研究和工业，例如蛋白质的改进，生物分子的合成，分子诊断和合成生物学。
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公开(公告)号：KR1020110060384A
公开(公告)日：2011-06-08
申请号：KR1020090116957
申请日：2009-11-30
Applicant: 한국화학연구원 , 충남대학교산학협력단
Abstract: PURPOSE: A Candida Antarctica-derived lipase B(CalB) protein variant is provided to increase enzyme activity and to be used in the industry. CONSTITUTION: A CalB protein variant is selected from a protein of amino acid sequences 2-8 in which 274-289th amino acid is partially substituted in Candida Antarctica-derived lipase B(CalB) protein. A gene encoding CalB protein variant sequence is denoted by sequence number selected from sequence numbers 10-16. A recombinant vector containing the gene is prepared by inserting the gen into pK7 vector. A transformed cell which produces CalB protein variant is obtained by transforming a host cell with the recombinant vector. The host cells are Pichia pastoris.
Abstract translation: 目的：提供来自南极假丝酵母的脂肪酶B（CalB）蛋白质变体以增加酶活性并用于该行业。 构成：CalB蛋白质变体选自氨基酸序列2-8的蛋白质，其中274-289位氨基酸被部分取代在南极假单胞菌来源的脂肪酶B（CalB）蛋白质中。 编码CalB蛋白变体序列的基因由选自序列号10-16的序列号表示。 通过将gen插入pK7载体中制备含有该基因的重组载体。 通过用重组载体转化宿主细胞获得产生CalB蛋白质变体的转化细胞。 宿主细胞是巴斯德毕赤酵母。
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Patent Agency Ranking
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公开(公告)号：KR102085636B1
公开(公告)日：2020-03-06
申请号：KR1020180164950
申请日：2018-12-19
Applicant: 충남대학교산학협력단
IPC: G01N33/548 , G01N33/543 , G01N33/68
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公开(公告)号：KR1020170014459A
公开(公告)日：2017-02-08
申请号：KR1020150107869
申请日：2015-07-30
Applicant: 충남대학교산학협력단 , 주식회사 파이안바이오테크놀로지
IPC: C12P21/00
Abstract: 본발명은무세포단백질합성방법에있어서, 무세포단백질합성반응액에포함된합성기질로서아미노산혼합물대신에효모추출물또는트립톤을이용하는것을특징으로하는무세포단백질합성방법에관한것이다. 본발명의무세포단백질합성방법은잘려진형태의단백질발현이감소하고, 전장(full-length)의단백질발현이증가하는효과가있는무세포단백질합성방법이다.
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公开(公告)号：KR101176802B1
公开(公告)日：2012-08-27
申请号：KR1020090116957
申请日：2009-11-30
Applicant: 한국화학연구원 , 충남대학교산학협력단
Abstract: 본 발명은 캔디다 안타르크티카(
Candida antarctica ) 유래의 리파제 B(CalB) 단백질 변이체에 관한 것으로서, 서열번호 1의 아미노산 서열로 표시되는 CalB 단백질의 274 위치 내지 289 위치의 아미노산 중 일부가 치환된, 서열번호 2 내지 8의 아미노산 서열로 표시되는 단백질로 이루어진 군에서 선택되는 본 발명에 따른 CalB 단백질 변이체는, 기존의 CalB 단백질보다 최대 35 배 증가된 효소 활성을 가지므로 리파제를 이용하는 산업에 유용하게 사용될 수 있다.
생명공학, 효소, 변이, 단백질, 활성-
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
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）。
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