Abstract:
PURPOSE: A NADPH quantitation using fluorescent protein his-mBFP is provided to quickly enhance fluorescence without oxygen and to measure a NADPH concentration in vivo. CONSTITUTION: A NADP(H) quantitation of an in vivo or in vitro sample comprises a step of measuring a fluorescence change induced by mBFP(metagenome-derived Blue Fluorescent Protein) coloration through mBFP containing an amino acid sequence of sequence number 3. The fluorescence change is measured by measuring a NADP(H) concentration or NADP(H)-dependent enzyme activity from increased or decreased fluorescence intensity. The enzyme is NADP(H)-dependent dehydrogenase or oxidoreductase. The quantitation further comprises a step of adding a surfactant during the reaction of the sample with mBFP. The surfactant is SDS, Na-deoxycholate, CTAB, DEDAB, or CHAPS. [Reference numerals] (AA) Escherichia coli with florescence on the basis of the increase of biological activity using polluted organic materials as nutrient
Abstract:
본 발명은 천연재료인 쪽 식물에 포함되어 있는 인디칸(indican)에 작용하여 염료와 의약품 원료, 생리활성 물질로 이용되는 인디루빈(indirubin)의 생산 방법에 관한 것으로 베타-글루코시다아제(β-glucosidase)와 이사틴 생성 효소를 코딩하는 유전자를 포함하는 형질전환 미생물로 인디루빈를 특이적으로 대량 생산할 수 있는 방법을 제공하는 것이다. 더욱 상세하게는, 본 발명은 인디루빈의 제조 과정 중 인독실의 C-2 위치에 선택적으로 산화활성을 나타내는 옥시게나아제의 발굴 및 그 개량방법에 관한 것으로, 생물정보학적 방법으로 GenBank 에 등록된 서열에서 관련 유전자를 탐색/발굴한 후 발현벡터에 삽입해 형질전환시킨 재조합 균주를 ampicillin 과 인디칸이 포함된 LB 고체배지에 도말해 붉은색을 띄는 클론을 확보하는 과정을 거쳐 선발된 기능성 옥시게나아제 및 그 제조방법을 제공하는 것이다. 또한 발굴된 옥시게나아제의 활성증대를 위해 무작위 돌연변이법으로 제작된 재조합 유전자를 글루코시다아제가 포함된 재조합 균주에 형질전환한 후 대조군에 비해 인디루빈 생성능이 증가된 유전자가 삽입된 클론을 확보하는 과정으로 개량된 유전자원을 이용하는 인디루빈 제조 방법을 제공하는 것이다. 옥시게나아제, 글루코시다아제, 인디칸, 인독실, 이사탄, 인디루빈
Abstract:
PURPOSE: A reduction method of indigo using yeast is provided to effectively reduce natural indigo and synthetic indigo in short time for improving the dye uptake of fabrics. CONSTITUTION: A reduction method of indigo uses a strain selected from the group consisting of saccharomyces cerevisiae, saccharomyces rosei, saccharomyces uvarum, saccharomyces chevalieri, hansenula spp, torulaspora delbrueckii, and kluyveromyces thermotolerans. A fabric dyeing method using natural indigo or synthetic indigo comprises a step of reducing an insoluble dye using the reduction method of the indigo, dipping fabrics into the reduced dye, and oxidizing the fabrics.
Abstract:
본 발명은 서열번호 2에 기재된 아미노산 서열을 코딩하는 에스테라아제(ESTL120P) 유전자 내에 다중 클로닝 위치(multiple cloning site, MCS)가 도입되어 있는 재조합 폴리뉴클레오티드에 관한 것으로, 상기 재조합 폴리뉴클레오티드를 인디케이터(indicator)로서 사용하여 외래 유전자를 포함하는 클론을 선별하는 단계를 포함하는 재조합 유전자 클로닝 벡터의 제조방법은 기존의 인디케이터(indicator)들 보다 저가의 기질을 이용하며 짧은 배양시간을 통해 빠르고 정확하게 재조합 균체의 선별이 가능하며, PCR 증폭 산물을 직접 클로닝할 수 있는 TA 클로닝 벡터와의 접목을 통해 상업적으로 판매되는 기존의 클로닝 벡터내의 인디케이터(indicator)들을 대체 할 수 있을 것으로 기대된다. 인디케이터, 에스테라아제, 다중 클로닝, 폴리뉴클레오티드
Abstract:
PURPOSE: A recombinant polynucleotide in which multiple cloning site is introduced into a beta glucosidase(cpGluT) gene is provided to accurately select recombinant strain. CONSTITUTION: A beta glucosidase(cpGluT) gene encodes an amino acid sequence of sequence number 2. The beta glucosidase gene comprises a base sequence of sequence number 1. A recombinant polynucleotide has multiple cloning site(MCS) in the beta glucosidase gene. A transgenic microorganism containing the recombinant polynucleotide is prepared by transforming a microorganism with a recombinant vector having the recombinant polynucleotide. The transgenic microorganism is deposited by deposit number KCTC 11500BP. A fusion protein contains the recombinant polynucleotide as a reporter.
Abstract:
PURPOSE: An esterase gene and recombinant polynucleotide are provided to replace an indicator in a conventional cloning vector through combining with TA cloning vector. CONSTITUTION: An esterase gene encodes an amino acid sequence of sequence number 2. The esterase gene comprises a sequence of sequence number 1. A recombinant polynucleotide has multiple cloning site in the esterase gene. The multiple cloning site is introduced one of sited from between 216th and 217th, 281th and 282th, and 306th and 307th in sequence number 2. The multi cloning site comprises a sequence of sequence number 12. The recombinant vector is TA cloning vector.
Abstract:
PURPOSE: A gene encoding an artificial restriction enzyme for constructing complete ORF library is provided to enhance the efficiency and screen the gene with low price in a short time. CONSTITUTION: A gene encoding an artificial restriction enzyme for constructing complete ORF library comprises: a cutting site of DNA or binding site of transcription factor; a protein binding site which recognizes transcription regulatory sequence or binds; and a protein which recognizes one selected among a promoter (-10, -35 site) of prokaryote and BRE, TATA, Inr or DPE of eukaryote.
Abstract:
PURPOSE: A transformed microorganism containing a gene encoding β-glucosidase or a method for producing indigo dye using the same is provided to ensure eco-friendly production and overcome problems of high cost and reproducibility. CONSTITUTION: A transformed microorganism containing a gene encoding β-glucosidase of sequences of the sequence number 1-8. The gene encodes β-glucosidase from a woad plant or its extract. A recombinant plasmid contains the gene and is expressed to maltose binding protein (MBP) fusion protein. A transformed microorganism E. coli XL1-BLUE[pTrc99A-glu(SM)] produces β-glucosidase from woad plant or its extract.