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1. 发明授权

KR100812110B1 징크 핑거 단백질과 원핵 생물의 전사 인자를 포함하는인공 전사 인자의 제조 및 이의 이용 有权
标题翻译：包含锌指蛋白和人工转录因子的人工转录因子的制备及其用途
公开(公告)号：KR100812110B1
公开(公告)日：2008-03-12
申请号：KR1020060103675
申请日：2006-10-24
申请人： 한국과학기술원
发明人： 김선창 , 이주영 , 성봉현 , 이준형 , 이상희 , 강귀현
IPC分类号： C07K14/435
CPC分类号： C07K14/4702 , C07K2319/81
摘要： An artificial transcription factor polypeptide is provided to control the activity of various prokaryotes by using a catabolite controlling protein of the prokaryotes as an effect domain, and induce various transformed E. coli(s) showing characteristics desired by a user through activation or inhibition of genes regardless of the action of the transcription factor inside of the E. coli by introducing the same thereinto, thereby obtaining the transformed E. coli with improved heat-resistance, low temperature resistance, high salinity resistance and growth speed. An artificial transcription factor polypeptide for activating or inhibiting gene expression artificially comprises 1 to 3 zinc finger domain(s) and a transcription factor of prokaryote, wherein the zinc finger domain is selected from the group consisting of zinc finger domains coded by nucleic acid sequences of SEQ ID : NOs. 13(Z1), 15(Z2), 17(Z3), 19(Z4), 21(Z5), 23(Z6), 25(Z7), 27(Z8), 29(Z9), 31(Z10), 33(Z11), 35(Z12), 37(Z13), 39(Z14), 41(Z15), 43(Z16), 45(Z17), 47(Z18), 49(Z19), 51(Z20), 53(Z21), 55(Z22), 57(Z23), 59(Z24), 61(Z25) and 63(Z26), and the transcription factor is selected from the group consisting of a wild type CRP(catabolite regulatory protein)(CRPW, residue 1-209) coded by SEQ ID : NO. 1, a CRP Del 137(residue 137-90) coded by SEQ ID : NO. 3 and a CRP Del 180(residue 1-180) coded by SEQ ID : NO. 5. A transformed E. coli is prepared by introducing the artificial transcription factor into the E. coli and has the heat-resistance and the resistance against the growth speed improvement, low temperature or high salt.
摘要翻译：提供人工转录因子多肽，以通过使用原核生物的分解代谢物控制蛋白作为效应结构域来控制各种原核生物的活性，并通过激活或抑制基因诱导显示用户期望的特征的各种转化的大肠杆菌不管大肠杆菌内部的转录因子的作用如何，通过引入其中，从而获得具有改善的耐热性，耐低温性，高耐盐性和生长速度的转化的大肠杆菌。用于人工激活或抑制基因表达的人工转录因子多肽包含1至3个锌指结构域和原核生物的转录因子，其中锌指结构域选自由锌指结构域编码的锌指结构域，所述锌指结构域由 SEQ ID：NO。 13（Z1），15（Z2），17（Z3），19（Z4），21（Z5），23（Z6），25（Z7），27（Z8），29（Z9），31（Z10） 33（Z11），35（Z12），37（Z13），39（Z14），41（Z15），43（Z16），45（Z17），47（Z18），49（Z19） 53（Z21），55（Z22），57（Z23），59（Z24），61（Z25）和63（Z26），转录因子选自野生型CRP（分解代谢调节蛋白）（CRPW，残基1-209）由SEQ ID NO： 1，编码SEQ ID NO：1的CRP Del 137（残基137-90）。 3和由SEQ ID NO：3编码的CRP Del 180（残基1-180）。通过将人工转录因子引入大肠杆菌中制备转化的大肠杆菌，并且具有耐热性和抗生长速度改善的抗性，低温或高盐。

2. 发明授权

KR100958096B1 염색체 특정 부위 제거용 재조합 벡터 및 이를 이용한미생물 내 염색체 특정 부위의 제거방법 有权
标题翻译：用于去除染色体的特定区域的重组载体和使用该染色体的微生物中特异性染色体区域的清除方法
公开(公告)号：KR100958096B1
公开(公告)日：2010-05-14
申请号：KR1020080012377
申请日：2008-02-11
申请人： 한국과학기술원
发明人： 김선창 , 강귀현 , 유병조 , 이준형 , 성봉현 , 이충훈 , 이상희 , 이주영 , 박명근
IPC分类号： C12N15/66 , C12N15/64 , C12N15/09
CPC分类号： C12N15/102 , C12N15/70 , C12N15/902 , C12N2800/80 , C12N2830/002
摘要： 본 발명은 염색체 특정 부위 제거용 재조합 벡터 및 이를 이용한 미생물 내 염색체 특정 부위의 제거방법에 관한 것으로, 더욱 상세하게는, 아라비노즈에 의해 유도되는 프로모터; 람다(λ)-레드(red) 재조합에 관여하는 단백질을 코딩하는 유전자; 람노즈에 의해 유도되는 프로모터; 및 I-
Sce I 제한효소를 코딩하는 유전자;를 포함하는, 염색체 특정 부위 제거용 재조합 벡터로, 기존 방법에 비해 보다 간편하고 빠르게, 선별마커를 남기지 않고, 미생물 내 특정 유전자를 연속적으로 제거할 수 있다.
유전자 제거, 람다-레드 재조합 (λ-red recombination), 람노즈, 아라비노즈, 유전체 재조합

3. 发明公开

KR1020090086870A 염색체 특정 부위 제거용 재조합 벡터 및 이를 이용한미생물 내 염색체 특정 부위의 제거방법 有权
标题翻译：用于去除染色体的特定区域的重组载体和使用该染色体的微生物中特异性染色体区域的清除方法
公开(公告)号：KR1020090086870A
公开(公告)日：2009-08-14
申请号：KR1020080012377
申请日：2008-02-11
申请人： 한국과학기술원
发明人： 김선창 , 강귀현 , 유병조 , 이준형 , 성봉현 , 이충훈 , 이상희 , 이주영 , 박명근
IPC分类号： C12N15/66 , C12N15/64 , C12N15/09
CPC分类号： C12N15/102 , C12N15/70 , C12N15/902 , C12N2800/80 , C12N2830/002
摘要： A recombinant vector for removing a specific site of chromosome is provided to sequencially and simply remove the specific gene site in a microorganism at once. A recombinant vector for removing a specific site of chromosome comprises a promoter (Para) which is induced by arabinose; a gene which encodes a protein related to lamda-red recombination; and promoter which is induced by a rhamnose. A method for removing the specific site of the chromosome using the recombinant vector comprises: a step of preparing a linear DNA fragment having homologous site A and B which is related to lamda-red recombination, I-SceI restriction enzyme cutting site, and homologous site C which is related to homologous recombination for removing selection marker; a step of introducing linear DNA fragement in transformed microorganism and replacing at specific site of microorganism chromosome; and a step of expressing I-SceI restriction enzyme and removing selection marker.
摘要翻译：提供了一种用于去除染色体特异性位点的重组载体，以便一次性地顺序并简单地除去微生物中的特异性基因位点。用于去除染色体特异位点的重组载体包含由阿拉伯糖诱导的启动子（Para）; 编码与lamda-red重组相关的蛋白质的基因; 和由鼠李糖诱导的启动子。使用重组载体去除染色体的特定位点的方法包括：制备与lamda-red重组，I-SceI限制酶切割位点和同源位点相关的具有同源位点A和B的线性DNA片段的步骤 C与同源重组相关，用于去除选择标记; 在转化的微生物中引入线性DNA分解并在微生物染色体的特定部位置换的步骤; 以及表达I-SceI限制酶并除去选择标记的步骤。

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