인체 α 1 - antitrypsin cDNA 의 분리 및 E . Coli 내에서의 발현

이상철,유명희 ( Sang Chul Lee,Myeong Hee Yu ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.2

Human α1-AT cDNA was screened from human liver cDNA library, by use of oligonucleotide probes. DNA insert of 1.3 kbp from a positive clone was subcloned into pUC9 plasmid and was analyzed by restriction enzyme digestion and partial DNA sequencing. The obtained gene was identical to human αl-AT cDNA reported previously. In order to express αl-AT cDNA under regulation of tac promoter, expression vector pT-AT was constructed. E. coli JM109 cells transformed with pT-AT showed αl-AT activity upon induction of recombinant protein synthesis. The recombinant αl-AT was immunologically identical to authentic αl-AT that was purified from human blood. When the recombinant cell lysates were analyzed on SDS-polyacrylamide gel electrophoresis, the expressed α1-AT was not detected by staining with Coomassie Brilliant Blue. However Western blotting of the same gel, using rabbit IgG against αl-AT and peroxidase-conjugated goat anti-rabbit IgG antibody, showed the existence of αl-AT at the expected molecular weight region. Molecular weight of recombinant αl-AT was slightly lower than that of authentic αl-AT; which presumably was due to lack of glycosylation inside E. coli cells.

고속 열차용 대형 알루미늄 부품의 GAP 대응 로봇 자동화 용접 기술

이상철,Lee, Sang Chul 대한용접접합학회 2013 대한용접·접합학회지 Vol.31 No.1

Robotized MIG welding of large aluminium components for high speed trains is state-of-the-art. The implementation of online laser cameras enables seam tracking and adoptive modification of welding parameters. A constant fill is achieved regardless of the gap tolerances. Friction Stir Welding has been introduced to the market as a reliable and fast joining technology. The advantages of high welding speeds and the elimination of arc light, fumes and liquefaction in the welding spot lead to economical realisation of heavy-duty gantry systems. FSW robots offer a high flexibility with regard to welding of curved parts, and can be equipped with laser cameras for exact joint tracking.

인체 ${\alpha}1$-antitrypsin cDNA의 분리 및 E.coli 내에서의 발현

이상철,유명희,Lee, Sang-Chul,Yu, Myeong-Hee 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.2

인체 간 cDNA library로부터 올리고 뉴클레오티드 프로브를 이용하여 ${\alpha}1$-antitrypsin (${\alpha}1$-AT) cDNA를 지닌 파지 클론을 스크린하였다. 선별된 클론에는 1.3 k bp의 DNA가 삽입되어 있었으며 이를 pUC9 벡터에 다시 클론닝하여 분석한 결과 이미 보고된 ${\alpha}1$-AT cDNA 서열과 일치함을 알 수 있었다. ${\alpha}1$-AT cDNA를 E. coli에서 발현시키기 위해 택 (tac) 프로모터에 의해 전사가 활성화되도록 발현벡터 pT-AT를 제조하였다. 플라스미드 pT-AT에 의해 형질전환된 E. coli JM109 균주에서 재조합 단백질의 발현을 유도한 결과 ${\alpha}1$-AT의 활성이 검증되었으며, 발현된 ${\alpha}1$-AT는 사람 피에서 정제한 ${\alpha}1$-AT와 그 면역학적 특성이 동일함을 알 수 있었다. 또한 같은 세포 추출액을 SDS-polyacrylamide 젤 전기영동법으로 분석한 결과 Coomassie Brilliant Blue로 단백질 밴드를 염색했을 때는 발현된 ${\alpha}1$-AT를 검증할 수 없었으나, ${\alpha}1$-AT에 대한 rabbit IgG 항체와 펄옥시다제가 연결된 anti-rabbit IgG goat 항체를 사용하여 웨스턴 블릿을 한 결과 ${\alpha}1$-AT 밴드가 기대했던 분자량 위치에서 확인되었다. 이와 같이 생산된 재조합 ${\alpha}1$-AT는 본래의 ${\alpha}1$-AT보다 분자량이 작은 것으로 나타났는데 이는 E. coli 세포내에서 발현된 ${\alpha}1$-AT는 glycosylation이 안된 형태의 것이기 때문일 것이다. Human ${\alpha}1$-AT cDNA was screened from human liver cDNA library, by use of oligonucleotide probes. DNA insert of 1.3 kbp from a positive clone was subcloned into pUC9 plasmid and was analyzed by restriction enzyme digestion and partial DNA sequencing. The obtained gene was identical to human ${\alpha}1$-AT cDNA reported previously. In order to express ${\alpha}1$-AT cDNA under regulation of tac promoter, expression vector pT-AT was constructed. E. coli JM109 cells transformed with pT-AT showed ${\alpha}1$-AT activity upon induction of recombinant protein synthesis. The recombinant ${\alpha}1$-AT was immunologically identical to authentic ${\alpha}1$-AT that was purified from human blood. When the recombinant cell lysates were analyzed on SDS-polyacrylamide gel electrophoresis, the expressed ${\alpha}1$-AT was not detected by staining with Coomassie Brilliant Blue. However Western blotting of the same gel, using rabbit IgG against ${\alpha}1$-AT and peroxidase-conjugated goat anti-rabbit IgG antibody, showed the existence of ${\alpha}1$-AT at the expected molecular weight region. Molecular weight of recombinant ${\alpha}1$-AT was slightly lower than that of authentic ${\alpha}1$-AT; which presumably was due to lack of glycosylation inside E. coli cells.

염화나트륨 농도가 느타리 자실체의 아미노산과 향기성분에 미치는 영향

이상철,편하영,박윤진,오태석,장명준,Lee, Sang-Chul,Pyeon, Ha-Young,Park, Youn-Jin,Oh, Tae-Seok,Jang, Myoung-Jun 한국버섯학회 2021 한국버섯학회지 Vol.19 No.4

본 연구는 염화나트륨에 의한 비생물적 스트레스가 생육과정 중의 느타리에게 미치는 영향을 자실체의 생육특성, 그리고 구성아미노산함량과 향기패턴 분석을 통해 확인하였다. 농도별로 염화나트륨을 처리한 톱밥배지에서 수확한 느타리 자실체의 생육특성을 조사한 결과, 처리구별 수량은 무처리구 대비 염화나트륨 0.5%처리구에서의 값은 비슷했으며, 염화나트륨 처리 농도가 1.0%에서 2.0%으로 증가하면서 수량이 확연히 감소하였다. 자실체의 구성아미노산함량 분석결과, 무처리구 대비 모든 염화나트륨 처리구에서 glutamic acid과 proline을 제외한 aspartate, threonine, serine, glycine, alanine, methionine, valine, isoleucine, leucine, phenylalanine, tyrosine, lysine, histidine, arginine의 14종 구성아미노산의 함량이 낮음을 확인하였다. 자실체의 향기패턴 분석결과, 염화나트륨 처리구에서 버섯고유의 향을 나타내는 octane compound로 추정되는 물질의 intensity가 감소하였음을 chromatography상에서 확인하였다. We investigated the effect of sodium chloride-associated abiotic stress on the development of Pleurotus ostreatus. We examined the growth characteristics of fruiting bodies, constituent amino acids, and fragrance pattern to determine the effect of culturing Pleurotus ostreatus on a sawdust substrate supplemented with sodium chloride in a dose-dependent manner. Pleurotus ostreatus fruiting bodies exhibited an increasing tendency towards augmented yields when grown in the presence of 0.5% sodium chloride as compared with that grown in the control group. However, increasing the supplementation of sodium chloride from 1.0 % to 2.0% resulted in significantly decreased yields of Pleurotus ostreatus fruiting bodies in these groups as compared with that in control groups. Further assessment revealed the presence of 14 types of amino acids in the fruiting bodies, including aspartate, threonine, serine, glycine, alanine, methionine, valine, isoleucine, leucine, phenylalanine, tyrosine, lysine, histidine, and arginine, at lower levels in all the sodium chloride-treated groups than in the control group; except for glutamic acid and proline. Similarly, fragrance pattern analysis of the Pleurotus ostreatus fruiting body by chromatography confirmed that the intensity of the substances presumed to be octane compounds, to which the unique flavor of mushrooms is attributed, was lower in all the sodium chloride-treated groups than in the control group.

치자(梔子)가 JNK와 NF-kB를 통한 항산화와 항염증의 대사과정에 미치는 영향

이상철,심성용,김연섭,Lee, Sang-Chul,Sim, Sung-Yong,Kim, Youn-Sub 대한한방안이비인후피부과학회 2016 한방안이비인후피부과학회지 Vol.29 No.2

Objectives : The purpose of this study is to observe the effects of Gardeniae Fuctus(GF) on the metabolic process of antioxidant and anti-inflammation. Methods : 4-HNE was injected into PC-12 cell to cause oxidative stress-induced inflammatory response, and then a western blot was conducted to observe the expression of Nuclear Factor-kB (NF-kB) and c-Jun N-terminal kinase (JNK) protein that are important factors involved with inflammation. Results : 1. The Gardeniae Fuctus water extract 50 ㎍ and 100 ㎍ significantly suppressed the increase in JNK protein expression in PC-12 cell. 2. The Gardeniae Fuctus water extract 50 ㎍, 100 ㎍ and 200 ㎍ significantly suppressed the increase in NF-kB protein expression in PC-12 cell. Conclusion : These results suggest that the Gardeniae Fuctus water extract has antioxidative and anti-inflammatory activity through suppressing the activity of JNK and NF-kB.

정보통신부 장관 취임사

이상철,Lee, Sang-Chul 한국정보통신집흥협회 2002 정보화사회 Vol.157 No.-

전산화단층사진을 이용한 타액선의 정량분석에 관한 연구

이상철,이삼선,허민석,최순철,박태원,유동수,Lee Sang-Chul,Lee Sam-Sun,Heo Min-Suk,Choi Soon-Chul,Park Tae-Won,You Dong-Soo 대한영상치의학회 1999 Imaging Science in Dentistry Vol.29 No.1

Purpose: The purpose of this study was to calculate the size and CT number of both normal parotid and submandibular gland. and evaluate their relation to sex, age and obesity using computed tomography. Materials and Methods: The computed tomography was performed parallel to the Frankfurt plane in 46 subjects with healthy salivary gland. The subjects were divided into the three groups (young, middle. old) according to their ages. The size of salivary gland was determined as maximum cross-sectional area and the CT number of salivary gland was determined as the mean CT number of three ROI's. The body mass index was calculated from weight and height. Results: The mean maximum cross-sectional area was 7.79(±1.25)cm² on parotid gland and 4.12(±0.83) cm² on submandibular gland. The mean CT number was -4.43(±23.87) HU on parotid gland and 50.01(±15.63) HU on submandibular gland. There was decreasing pattern of the maximum cross-sectional area of submandibular gland and the CT number of both parotid and submandibular gland according to age(p<0.05). As the body mass index increased. the maximum cross-sectional area of parotid gland increased and CT number of both parotid and submandibular gland decreased(p<0.05). The maximum cross-sectional area of submandibular gland in male was larger than that in female(p<0.05). As the maximum cross-sectional area and CT number of left salivary gland increased. those of right gland increased(p<0.05). Conclusion : Intra-individual differences in salivary gland size and CT number is considered in the age and individual obesity.

Rhizobium trifolii Malonyl-CoA Synthetase의 촉매반응에 관여하는 아미노산 Residue의 화학변형

이상철,김유삼,Lee, Sang-Chul,Kim, Yu-Sam 생화학분자생물학회 1993 한국생화학회지 Vol.26 No.3

Rhizobium tnifolii에서 정제한 malonyl-CoA synthetase가 2,3-butanedione과 diethylpyro-carbonate(DEP)에 의해 불활성화 되었다. 각 불활성화 반응은 pseudo first-order kinetics으로 나타났다. 2,3-Butanedion에 의한 효소의 불활성화 반응의 2차 속도상수는 pH 8.0, $30^{\circ}C$에서 $17.4M^{-1}{\cdot}min^{-1}$이었다. DEP에 의해 효소가 빠르게 불활성화되었으며 반응의 2차속도상수는 pH 7.1, $30^{\circ}C$ 에서 $780M^{-1}{\cdot}min^{-1}$이었다. 각 시약과 반응차수는 1.06과 1.03이였다. ATP가 위의 2가지 시약에 의한 효소의 불활성화를 저해하였다. 이러한 결과들은 malonyl-CoA synthetase의 ATP 결합부위나 그 주위에 효소활성에 필수적인 arginine residue와 histidyl residue가 존재함을 시사한다. Malonyl-CoA synthetase from Rhizobium trifolii was inactivated by 2,3-butanedione and diethylpyrocarbonate (DEP). In each case, inactivation followed pseudo first-order kinetics. The second-order rate constant for the inactivation by 2,3-butanedione was $17.4M^{-1}{\cdot}min^{-1}$ at pH 8.0, $30^{\circ}C$. DEP rapidly inactivated the enzyme with the second-order rate constant, $780M^{-1}{\cdot}min^{-1}$ at pH 7.1, $30^{\circ}C$. The reaction order of inactivation with respect to reagents were 1.06 and 1.03 respectively. The substrate, ATP, afforded the protection against the inactivation of the enzyme caused by the reagent, respectively. These results indicate that arginine and histidyl residues essential for the enzyme activity are located at or near the ATP binding region of the active site.

Determinants of Effecting Customer Loyalty : Comparison among Korean, Japanese and Chinese Online Game Market

이상철,상균영,구자철,서영호,Lee, Sang-Chul,Xiang, Jun-Yong,Gu, Ja-Chul,Suh, Yung-Ho The Korean Operations Research and Management Scie 2006 經營 科學 Vol.23 No.2

The purposes of this research are to identify causalities among flow and customer loyalty In Chinese online games, and to identify the factors by which flow are influenced. This research tests the model with Chinese on-line game users and compare this result with Korea and Japanese results which were conducted by Lee's research. These implications are thought to be helpful for Korean online game companies to understand the Chinese online game user and to develop the penetration strategies. The results indicated that significant Path coefficients to flow were the convenience of operator, the provision of information, the reality of design. The results indicated that significant path coefficients to customer loyalty were the involvement of virtual community and flow. The involvement of virtual community to flow was not significant but to customer loyalty was significant. The provision of information was negatively influenced on flow. The result of comparison indicated that the path coefficients were different among nations. Korea online game companies need to develop the indigenized online game and to Provide the information to their Chinese partner correctly and quickly.

Pyridoxal-5'-Phosphate에 의한 Rhizobium trifolii Malonyl-CoA Synthetase의 화학변형

이상철,김유삼,Lee, Sang-Chul,Kim, Yu-Sam 생화학분자생물학회 1993 한국생화학회지 Vol.26 No.3

Pyridoxal-5’-phosphate가 Rhizobium tifolii에서 정제한 malonyl-CoA synthetase를 가역적으로 불활성화 하였다. 반응의 2차 속도상수는 pH 7.0, $30^{\circ}C$에서 $2975 M^{-1}{\cdot}min^{-1}$이었다. PLP에 의해 불활성화된 효소는 완충용액으로 희석하였을 때 가역적으로 활성을 회복하였으나, $NaBH_4$로 환원 후 희석한 효소는 활성을 회복하지 못하였다. PLP와 반웅 후 $NaBH_4$로 환원한 효소액은 325 nm에서 톡징적인 Schiff base의 홉광도 변화를 보였다. ATP가 PLP에 의한 효소의 불활성화를 저해하였다. 이러한 결과들은 malonyl-CoA synthetase의 ATP 결합부위나 그 주위에 효소활성에 필수적인 amine group이 존재함을 시사한다. Pyridoxal-5'-phosphate inactivated malonyl-CoA synthetase from Rhizobium trifolii reversibly. The second-order rate constant, $K_i$ was $2975 M^{-1}{\cdot}min^{-1} $ at pH 7.0, $30^{\circ}C$. Inactivation of malonyl-CoA synthetase by PLP was reversed by dilution but not after reduction with $NaBH_4$. The spectra of malonyl-CoA synthetase, inactvated by PLP and reduced by $NaBH_4$, showed a characteristic Schiff base absorption change at 325 nm. ATP protected the enzyme from inactivation by PLP. These studies indicated that an essential amine is located at or near ATP binding region of the enzyme active site.