• 한국미생물생명공학회:학술대회논문집
• /
• 한국미생물생명공학회 1986년도 추계학술대회
• /
• pp.516.2-516
• /
• 1986

DNA subfragments, sopA, sopB, and sopC supporting stable maintenance of an oriC plasmid, were derived from mini-F plasmid DNA (EcoRI restriction fragment, f5) after digestion with restriction endonucleases, and cloned in vector plasmid pBR322. The recombinant plasmid obtained were introduced into E. coli KY7231 and E. coli CSR603, and proteins specified by the mini-F fragments were analysed by SDS-polyacrylamide gel electrophoresis. Two proteins encoded by the F fragments were detected, having molecular weights of 41,000 and 37.000. The sopA protein (41K) encoded by a plasmid pXX288 was observed in the cytoplasm, whereas the sopB protein (37K) encoded by a plasmid pXX157 was in the membrane fraction. There was no novel protein band detected in the cell with a plasmid pXX300, which contained sopC fragment. Gene products of a plasmid pXX167, which is comprised of sopA, sopB, and sopC, were not detectable. Fluorography after one and two dimensional gel electrophoresis of the lysates showed that these two proteins were overproduced in the cells which were allowed to incorporate radioactive amino acid after plasmid amplification by chloramphenicol treatment. The isoelectric points of the sopA and sepB proteins were 6.6 and 7.0, respectively.

• The Korean Journal of Physiology
• /
• 제25권2호
• /
• pp.115-123
• /
• 1991

Molecular association of glucose transporters with the other proteins in the plasma membrane was assessed by gel electrophoresis and immunoblot techniques. Approximately $31.5{\pm}5.1%$ of GLUT-4, $64.8{\pm}2.7%$ of clathrin, 48.7% of total protein in the plasma membrane (PM) were found insoluble upon extraction with 1% Tx-100. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the Tx-100 insoluble PM fraction contained about 4 major polypeptides with apparent molecular weight of above 200, 100-120, 80 and 30-35 KDa that were readily removed upon wash with a high pH buffer which is known to remove clathrin and 0.5 M Tris-buffer which is known to remove assembly proteins (AP). Immunoblotting of GLUT4 and clathrin against specific antibodies showed that GLUT-4 and clathrin were co-solubilized up to 84.6% and 82.7% respectively by wash with a high pH buffer and 1% Tx-100. When the membrane was pre-washed with a high pH buffer and 0.5 M Tris solution, GLUT4 and clathrin were not solubilized further suggesting that GLUT4 molecules are in molecular association with clathrin, AP and/or other extrinsic membrane proteins in plasma membrane and the formation of clathrin-coated structures might be involved in insulin stimulated glucose transporter translocation mechanism.

• 한국미생물·생명공학회지
• /
• 제22권1호
• /
• pp.73-79
• /
• 1994

The optimum cultural temperature and time for the pullulanase production by Bacillus cereus subsp. mycoides were 35$\circ $C and 48 hrs, respectively. The addition of egg albumin and casein to the basal medium increased the enzyme production. The enzyme was purified by ammonium sulfate fractionation and DEAE-cellulose column chromatography. specific activity of the purified enzyme was 82.37 U/mg protein and yield of theenzyume activity was 62.1%. The purified enzuyme showed a single band on ployacrylamide disc gel electrophoresis and its molecular weight was estimated to be 66.,000 by SDS-polyacrylamide disc gel electrophoresis. The isoelcular point for the purified enzyme was pH 5.0. The optimum temperature and pH were 50$\circ $C and pH 6.5, respectively. The purified enzyme was stable below 40$\circ $C and in the pH range of 6.5~10.0 The pullulanase activity was greatly inhited by Ag$^{+}$, Hg$^{2+}$ and EDTA, and its heat stability was increased by the addition of Ca$^{2+}$. The tydrolysis product with the enzyme on pullulan was maltotriose.

• 한국잠사곤충학회지
• /
• 제31권2호
• /
• pp.72-81
• /
• 1989

천잠의 알에서 난황 주단백질인 vitellin을 분리·정제하고 가잠 및 상잠의 vitellin과 첨잠 nitellin과의 면역학적 특성을 비교하였으며 천잠의 배자발생에 따른 난내 vitellin의 변동을 조사하였다. 1. 전잠의 체액 vitellogenin은 유충기의 토사말기부터 검출되었다. 2. 점잠의 vitellinm은 가잠 및 상잠의 vitellin과 저기영동상의 이동도는 같았으나 면역학적인 특성에 있어서는 비동질성이었다. 3. 천잠 vitellin은 함량은 전체 가용성 난단백질의 46.0%였으며, 배자발생에 따른 vitellin의 변동은 산난후 8일까지는 같은 수준이었나, 10일 이후부터는 감소되는 경향을 보이다가 5개월된 난과 17개월된 난에서는 상당히 미량만이 검출되었다.

• 한국응용곤충학회지
• /
• 제29권3호
• /
• pp.170-175
• /
• 1990

점박이응애(Tetranychus urticae Koch)의 살비제저항성 기작을 구명하기 위한 기초시험으로서, 유기인계 carbophenothion과 ethion, 유기염소계 dicofol, 유기주석계 cyhexatin 및 합성 pyrethroid계 biphenthrin으로 누대도태한 각 저항성계통과 감수성계통을 공시하여, esterase isozyme의 영동대 차이점 (polyacrylamide gel electrophoresis)을 비교한 결과, carbophenothion 저항성계통은 감수성계통에서 나타나지 않은 Est. 1, Est. 3이 검출되었고, ethion과 cyhexatin 저항성계통에서는 각각 Est. 3이, dicofol 저항성계통에서는 Est. 1, Est. 3, Est. 7이, biphenthrin 저항성계통은 Est. 3, Est. 7이 검출되었다. 이러한 영동대와 기질분해량의 차이점으로 미루어 보아 공시살비제들의 저항성발달에는 esterase가 관여하는 것으로 나타났다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제17권3호
• /
• pp.420-422
• /
• 2004

This study was to demonstrate a rapid novel method for detection of adulterated cow milk in goat milk using modified staining protocol after native polyacrylamide gel electrophoresis (PAGE). Samples of cow milk and goat milk containing 0, 0.5, 1.0 and 2.0% (v/v) of cow milk were analyzed. Low levels of cow milk mixed in goat milk were identified by the presence of higher mobility of $\beta$-lactoglobulin A ($\beta$-Lg A) in cow milk. By mini-gel electrophoresis, a distinguishable protein profile was visualized in 25 min using the modified Coomassie blue staining solution, in which methanol (50%) was replaced with ethanol (20%) and the concentrations of Coomassie blue and acetic acid were reduced from 2 to 0.13% and 10 to 5%, respectively. To visualize the milk proteins, gels in the staining solution were water-bathed in boiling water for 5 min and then cooled down immediately for 3-5 min. The sensitivity of this method is relatively high, allowing examination of 1% cow milk in goat milk. The procedure presented here is also very cost-effective due to less reagents needed. This simplified method would be useful and applicable to dairy industry for routine examination of goat milk.

• 한국동물학회지
• /
• 제28권2호
• /
• pp.61-70
• /
• 1985

斧足綱 數種의 蛋白質을 電氣泳動한 結果, 새꼬막과 피조개의 血漿의 蛋白質 패턴은 低分子 蛋白質에서 染色되는 强度가 약간 差異가 있으나 거의 同一하였고, 全血球의 蛋白質 패턴의 差異는 17,800 dalton의 band에서 나타났다. 그렇지만 위의 두 種의 筋內의 단백질 패턴은 거의 비슷해서 10,000$\\sim$100,000 dalton의 分子量 範圍內에서는 差異를 알아 내기가 매우 어려웠다. 그런데 대합과 비늘백합은 類緣關係가 있는 蛋白質 패턴이 많이 나타났으며 재첩은 대합이나 비늘백합과 같은 이들 두 種과 대칭이의 中間 程度에 位置하는 蛋白質 패턴을 나타내고 있었다. 본 연구에서는 Bivalvia가 함께 다 갖는 6개의 蛋白質 band들과, 꼬막과 새꼬막에서만 나타나는 4개의 特有한 蛋白質 band가 存在함이 發見되었으며 Eulamellibranchia目의 4종에 있어서 다 나타나는 2개의 protein band들 과, 대합과 비늘백합에서만 나타나는 23,000 dalton의 特有한 band도 發見되었다. 그리고 各 種마다 가지는 特有한 蛋白質 패턴의 分子量을 測定하여 比較하였다.

• BMB Reports
• /
• 제32권6호
• /
• pp.579-584
• /
• 1999

Mantis egg fibrolase (MEF-3) was purified from the egg cases of Tenodera sinensis using ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, DEAE Affi-Gel blue gel affinity chromatogragphy, and MONO-Q anion-exchange chromatography. This protease had a molecular weight of 35,600 Da as determined by SDS-polyacrylamide gel electrophoresis under reducing conditions and its isoelectric point was 6.0. The N-terminal amino acids sequence was Ala-Thr-Gln-Asp-Asp-Ala-Pro-Pro-Gly-Leu-Ala-Arg-Arg. This sequence was 80% homologous to the serine protease from Tritirachium album. MEF-3 readily digested the ${\alpha}$-and ${\beta}$-chains of fibrinogen and more slowly the ${\gamma}$-chains. It showed strong proteolytic and fibrinolytic activities. Phenylmethanesulfonyl fluoride and chymostatin inhibited its proteolytic activity, while EDTA, EGTA, cysteine, ${\beta}$-mercaptoethanol, elastinal, tosyl-lysine chloromethylketone, and tosyl-amido-2-phenylethyl chloromethyl ketone did not affect its proteolytic activity. Among the chromogenic protease substrates, the most sensitive one to the hydrolysis of MEF-3 was benzoyl-Phe-Val-Arg-p-nitroanilide. Based on these experimental results, we speculated that MEF-3 is a serine protease with a strong fibrin(ogen)olytic activity.

• 한국미생물·생명공학회지
• /
• 제17권5호
• /
• pp.440-446
• /
• 1989

Penicillium sp. CB-20의 polygalacturonase 생성을 위한 최적조건은 탄소원으로 펙틴을 사용하여 16 시간 배양시 최대 활성을 나타내었으며, Sephadex G-25, G-75 및 G-150을 사용한 gel filtration과 DEAE-cellulose, DEAE-Sephadex A-50에 의 한 ion exchange chromatography를 통하여 이 효소를 약 30배 가량 정제할 수 있었고, 수율은 2.31%였다. 정제효소는 polyacrylamide gel electrophoresis에 의하여 단일 밴드로 확인 되었으며, 분자량은 SDS PAGE에 의하여 21,000 정도로 측정되었다. 효소의 결정구조는 마름모꼴을 형성하고 있었으며 아미노산 조성은 17종류로써 glutamic acid, glycine, hlstidine의 함량이 비교적 많았다.

• Applied Biological Chemistry
• /
• 제27권4호
• /
• pp.271-277
• /
• 1984

Pseudomonas stutzeri IAM 12097이 생산(生産)하는 Exo-maltotetraohydrolase의 gel filtration에 의하여 추정(推定)된 분자량(分子量)은 60,000이었고, SDS-polyacrylamide gel electrophoresis에 의하여 추정(推定)된 분자량(分子量)은 63,000이었으며 등전점(等電點)은 PH 4.8이었다. 최적(最適)pH는 6.6, PH안정(安定)범위는 $6.0{\sim}10.5$, 최적온도(最適溫度)는 $45{\sim}50^{\circ}C,\;40^{\circ}C$이하(以下)에서는 안정(安定)하였으며, $55^{\circ}C$이상(以上)에서는 급격하게 불활성화(不活性化)되었다. 본효소(本酵素)는 $Ag^+,\; Hg^{++},\;I_2,\;{\beta}-cycoldextrin$에 의하여 완전(完全)히 저해(沮害)되었고 EDTA, ${\rho}-CMB$, IAA에 의하여 약간 저해(沮害)되었다. soluble starch, amylopectin, amylose에 대(對)한 Michaelis constant(Km)는 각각(各各) 7.70mg/ml, 6.17mg/ml, 5.56mg/ml이었다.