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11.发明授权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蛋白质表达水平的初始密码子的方法，以及用于调节重组蛋白质的表达和产生的方法
公开(公告)号：KR100961565B1
公开(公告)日：2010-06-07
申请号：KR1020080022217
申请日：2008-03-10
Applicant: 충남대학교산학협력단
CPC classification number: C12N15/67 , C12N15/102
Abstract: 본 발명은 유전자의 초기 코돈(initial codon), 즉 +2 내지 +13 코돈 중 하나 이상, 특히 +2 및 +3 코돈을 무작위로 변화시킨 후 이들을 세포 내로 도입하여 개별 유전자로 분리하고 콜로니 PCR에 의해 증폭한 후, 무세포 단백질 합성 시스템을 이용하여 각각의 유전자를 단백질로 발현시킴으로써, 목적하는 단백질 발현 수준을 제공하는 초기 코돈을 발굴하는 방법, 및 이를 이용한 재조합 단백질의 발현 조절 방법 및 생산 방법에 관한 것으로서, 단백질 생산에 있어 체계적이고 정확한 조절 수단이 될 수 있으며, 활성 단백질을 원하는 수준으로 고속으로 발현할 수 있다.
역단백질체학, 리버스 프로테오믹스, 초기 코돈, 고속 단백질 발현, 무세포 단백질 합성-
12.发明公开단백질 어레이의 동소 생성을 위한 지지체 및 유전자 어레이, 이로부터 얻어지는 단백질 어레이, 및 이의 제조 및 사용 방법 有权蛋白质阵列产生蛋白质的支持和基因组，获得的蛋白质阵列及其制备和使用方法
公开(公告)号：KR1020100052806A
公开(公告)日：2010-05-20
申请号：KR1020080111667
申请日：2008-11-11
Applicant: 충남대학교산학협력단
CPC classification number: B01J19/0046 , B01J2219/00387 , B01J2219/00527 , B01J2219/00596 , B01J2219/00605 , B01J2219/00612 , B01J2219/0063 , B01J2219/00637 , B01J2219/00641 , B01J2219/00659 , B01J2219/00722 , B01J2219/00727
Abstract: PURPOSE: A support and gene array for in situ generation of protein array is provided to quickly convert various kinds of DNA or mRNA to active protein of array form and enhance protein array analysis sensitivity. CONSTITUTION: A support for in situ generation of protein array contains a solid support and hydrogel which is coated on the solid support and comprise protein biosynthesis machinery. The hydrogel is agarose gel. The hydrogel is manufactured by spin coating in a film form. A gene array for in situ generation of protein array comprises the support for in situ generation of protein array and a genetic template spotted on the support.
Abstract translation: 目的：提供用于原位产生蛋白质阵列的支持和基因阵列，以将各种DNA或mRNA快速转换为阵列形式的活性蛋白，并增强蛋白质阵列分析的灵敏度。 构成：原位产生蛋白质阵列的支持物包含固体支持物和水凝胶，其被包被在固体支持物上并且包括蛋白质生物合成机制。 水凝胶是琼脂糖凝胶。 水凝胶通过以薄膜形式旋涂制造。 用于原位产生蛋白质阵列的基因阵列包括支持蛋白质阵列的原位产生和在支持物上发现的遗传模板。
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13.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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Patent Agency Ranking
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：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：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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