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권정숙,박종상 ( Jung Sook Kwon,Jong Sang Park ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.5
Glucose-6-phosphatase has been known as a multicomponent system and glucose-6-phosphatase was labeledd by incubating with [^(32)P]glucose-6-phosphate to be saturation concentration. Microsomal pellet was obtained by the method of differential ultracentrifugation and distrupted with sonication. To this microsomal protein mixture, [^(32)P]glucose-6-phosphate was added and incubated. Then the mixture was seperated with 10∼15% gradient PAGE and the gel was dried and exposed on the X-ray film. As a result, 60 kd band was obtained and this was considered as a glucose-6-phosphatase. The same result was obtained when [^(32)P]mannose-6-phosphate and [^(32)P]galactose-6-phosphate were used as substrates. 20% trichloro acetic acid. 8 M urea, and 10% SDS were used as a quenching system respectively and 60 kd band was still unchanged.
Steady-State Labeling of Hepatic Glucose-6-phosphotase
권정숙,박종상,Kwon, Jung-Sook,Park, Jong-Sang 생화학분자생물학회 1991 한국생화학회지 Vol.24 No.5
글루코즈-6-인산 가수분해 효소에 $^{32}P$로 표시된 글루코즈-6-인산을 과량 넣어 주어서 steady-state의 상태에서 글루코즈-6-인산 가수분해 효소를 표지시킨다. 쥐간에서 초고속 원심분리 방법으로 마이크로좀을 얻고 이 단백질 혼합물에$[^{32}P]$ 글루코즈-6-인산을 넣어 반응 후, SDS 젤에서 전기영동으로 분리, 건조하여 X-ray film으로 감광시킨 결과 60 kd의 한 밴드를 보았다. 한편, 만노즈-6-인산과 갈락토즈-6-인산을 기질로 사용해서도 똑같은 밴드를 보았다. 또한 단백질 변성 방법으로 8M 우레아를 쓰거나, 20% Trichloro-acetic acid 혹은 2% SDS를 사용해도 여전히 같은 위치에서 밴드가 나타난다. Glucose-6-phosphatase has been known as a multicomponent system and glucose-6-phosphatase was labeledd by incubating with $[^{32}P]$glucose-6-phosphate to be saturation concentration. Microsomal pellet was obtained by the method of differential ultracentrifugation and distrupted with sonication. To this microsomal protein mixture, $[^{32}P]$glucose-6-phosphate was added and incubated. Then the mixture was seperated with 10∼15% gradient PAGE and the gel was dried and exposed on the X-ray film. As a result, 60 kd band was obtained and this was considered as a glucose-6-phosphatase. The same result was obtained when $[^{32}P]$mannose-6-phosphate and $[^{32}P]$galactose-6-phosphate were used as substrates. 20% trichloro acetic acid. 8 M urea, and 10% SDS were used as a quenching system respectively and 60 kd band was still unchanged.
Cloning and Sequence Analysis of 3'-Terminal Region of Human Insulin Receptor Gene
이정화,박선희,박종상,Lee, Jung-Hwa,Park, Sun-Hee,Park, Jong-Sang Korean Society for Biochemistry and Molecular Biol 1990 한국생화학회지 Vol.23 No.3
사람 인슐린 수용체 유전자의 3'말단 부분의 5 kb를 클로닝하였다. 건조젤 하이브리디제이션을 이용하여 제한효소인 EcoRI으로 자른 사람 인슐린 수용체 유전자 중 4.5-5.5 kb를 확인하고 ${\lambda}gt$ 10벡터에 삽입시켰다. 이렇게 하여 만든 유전자 라이브러리를 카르복시 말단에 해당하는 합성 NDA로 스크린하여 표지를 내는 plague들을 pBR328에 재클로닝하였다. 부분적인 DNA의 염기서열 결정으로 클로닝한 DNA가 사람 인슐린 수용체임을 확인하였다. ExoIII nuclease로 DNA를 크기순서대로 자르는 방법을 이용하여 전사 말단 부위의 1.3 kb의 유전자 염기서열을 결정하였다. 염기서열 분석결과 인슐린 수용체 유전자는 poly(A)가 붙는 자리 앞쪽의 15 뉴클레오티드에 전통적인 AATAAA 염기가 아닌 AATATA 염기가 존재함을 알았는데 아마도 이것이 유전자 조절에 관여할 것으로 여겨진다. 또한 이 유전자는 mRNA 프로세서에 관여한다고 제안된 G/T 밀집 염기서열을 가지고 있음을 알았다. The 5 kb long 3'-terminal region of genomic DNA of Human Insulin Receptor(IR) gene was cloned. By the dry-gel hybridization, 4.5-5.5 kb long DNA of EcoRI cut human chromosomal DNA was identified and inserted into ${\lambda}gt$ 10 vector. Constructed subgenomic library was screened with synthetic 21 mer probe corresponding to the carboxy terminal. By the partial DNA sequencing about 200 bp in 5 kb genomic DNA, we found the sequence was the same as the published result. We sequenced about 1.3 kb transcription terminal region by the ExoIII deletion method. The result of the sequence analysis showed that the IR gene has AATATA sequence 15 nucleotide upstream of poly(A) site instead of the canonical AATAAA consensus sequence and this may play a role in gene regulation and that in poly(A) downstream region the IR gene has the T-rich, G/T cluster sequence, which was proposed as a element of mRNA processing.
Reduced Burst Release from ePTFE Grafts: A New Coating Method for Controlled Drug Release
Hye Yeong Nam,이병하,박종상,김대중,Hyun Jung Lim,Insu Baek,Sang Hun Park 대한화학회 2008 Bulletin of the Korean Chemical Society Vol.29 No.2
Hemodialysis graft coated with paclitaxel prevents stenosis; however, large initial burst release of paclitaxel causes many negative effects such as drug toxicity and inefficient drug loss. Therefore we developed and tested a novel coating method, double dipping, to provide controlled and sustained release of paclitaxel locally. Expanded polytetrafluoroethylene (ePTFE) grafts were dipped twice into a solution of several different paclitaxel concentrations. In vitro release tests of the double dipping method showed that early burst release could be somewhat retarded and followed by sustained release for a long time. We observed the effect of paclitaxel coating by double dipping in porcine model of arterio-venous (AV) grafts between the common carotid artery and the external jugular vein. 12 weeks after constructing AV grafts, cross sections of the graft venous anastomosis were obtained and analyzed. Paclitaxel coated ePTFE grafts by double dipping were observed to prevent neointimal hyperplasia and therefore reduced stenosis of the arteriovenous hemodialysis grafts, especially at the graft venous anastomosis sites. Our results demonstrate that second dipping of ePTFE graft, which was already coated once with paclitaxel, washes off the drug on a surface of the graft and affects the ratio of paclitaxel on the surface to that of the inner space, possibly by diffusion: thus the early burst of drug can be somewhat reduced.
枸杞子 突然變異 系統의 作物學的 特性과 RAPD로 본 遺傳的 變異
Bong Chun Lee(李鳳春),Tae Soon Kwak(郭泰淳),Jung Sang Park(朴琮祥),Kee Won Yu(庾基元) 한국육종학회 1997 한국육종학회지 Vol.29 No.4
To obtain the good quality, disease resistance and high yield of Boxthorn cultivar, seeds of cultivar in Chongyang native were treated with gamma rays of ⁶⁰Co 3KR and 6KR, and also the stem and seed of Yuseong 2 were treated with gamma ray of ⁶⁰Co 3KR, respectively. For mutant strains originated from different treatments, growth characteristics, yield and genetic difference using RAPD analysis were investigated. The mutations observed were changes in stem length, leaf area, numbers of branches and 100 seed weight, stem diameter, disease and pest resistance of mutant strains. Fruit setting and yield potential were abundant in mutated seeds and stem of Yuseong 2. On the other hand we have performed polymerase chain reactions on six chromosomal DNA using by forty random primers and followed by RAPD fragments. And also we selected ten primers which amplified to chromosomal DNAs and its PCR-DNA band size were below to 2.0 bp. To konw genetic similarity, we analysis polymorphisms of PCR-DNA bands by statistics, and its genetic difference distance were 0.7~1.0. The Boxthorn of mutant strains treated with Co⁶⁰ were discussed with morphological feature and RAPD-DNA bands.
대장상피세포에서 Interleukin-10 유전자 전달의 CXC 케모카인에 대한 억제 효과
이국래 ( Kook Lae Lee ),김찬규 ( Chan Gyoo Kim ),김병관 ( Byeong Gwan Kim ),장동경 ( Dong Kyung Chang ),이동호 ( Dong Ho Lee ),김주성 ( Joo Sung Kim ),정현채 ( Hyun Chae Jung ),이연 ( Yeon Lee ),박종상 ( Jong Sang Park ),송인성 ( 대한소화기학회 2003 대한소화기학회지 Vol.41 No.6
Background/Aims: Cytokine plays an important role in initiation and continuation of inflammatory bowel disease. However, cytokine protein has some limitation as a therapeutic tool because of low bioavailability, poor pharmacokinetics and chemical instability. Thus, we studied the effect of interleukin 10 (IL-10) gene transfection on murine colon cancer cell line by using non-viral gene carrier. Methods: Therapeutic gene and plasmid was pCAGGS mouse IL-10 and gene carriers were polyethyleneimine (PEI) and 3β[L-ornithinamide-carbamoyl] cholesterol (O-chol). After IL-10 gene transfection, we measured the level of IL-10 in supernatant of cultured CT-26 cells. The chemokine cytokine-induced neutrophil chemoattractant (KC) and macrophage inflammatory protein (MIP)-2, which were treated with lipopolysaccharide (LPS) or tumor necrosis factor-alpha (TNF-α), were measured after IL-10 gene transfection. Results: The IL-10 values were increased significantly by using PEI, but not by using O-chol. The KC and MIP-2 values were increased when LPS or TNF-α were treated. When PEI was used, KC and MIP-2 values increased by LPS or TNF-α were decreased. When O-chol was used, the KC values increased by TNF-α were decreased but those treated by LPS were not decreased, and the MIP-2 values were not decreased. Conclusions: After IL-10 gene transfection in colon cancer cell, IL-10 cytokine was efficiently expressed. The increased chemokine values by LPS or TNF-α were suppressed by IL-10 gene transfection, but which was not constant because of carrier efficiency. (Korean J Gastroenterol 2003;41:447-455)