Purification and Characterization of Hpa I endonuclease

윤호섭,강선철,유욱준,Yoon, Ho Sup,Kang, Sun Chul,Yoo, Ouk Joon 한국미생물 · 생명공학회 1985 한국미생물·생명공학회지 Vol.13 No.1

Hpa I endonuclease를 순수 정제하였다. 150g (wet weight)의 Haemophilus parainfluenzae로 부터 얻은 crude extract를 ammonium sulfate fractionation을 거친후 Heparin agarose, SP-sephadex, DEAE-sephadex, phosphocellulose 등 chromatography를 거쳐 최종적으로 0.2mg의 효소를 얻었다. Specific activity는 $2.2{\times}10^6units/mg$이었다. 얻어진 Hpa I endonuclease는 SDS-polyacrylamide gel에서 한개의 band를 보였고, 그 효소활성은 50mM NaCl과 pH 7.0과 7.5사이에서 가장 높았다. Hpa I endonuclease from Haemophilus parainfluenzae has been purified of homogeneity and its physical and ezymatic properties have been studied. For the purification of the enzyme, Heparin agarose, SP-sephadex C-25, DEAE-sephadex A-50 and phosphocellulose chromatography columns were used. The denatured and reduced form of the enzyme is a monomer of molecular weight of $30,000{\pm}1,000$ as judged by 10% polyacrylamide gel electrophoresis containing 0.1% sodium dodesyl sulfate. Hpa I endonuclease was maximally active at neutral pH (7.0 to 7.5) in the presence of 50 mM NaCl.

가막만 모자반 군락지 어류군집의 종조성 및 계절변동

윤호섭,안윤근,최상덕,Yoon, Ho-Seop,An, Yun-Keun,Choi, Sang-Duk 해양환경안전학회 2011 해양환경안전학회지 Vol.17 No.1

Species composition and seasonal variation of fish assemblages in Sargassum beds in Gamak Bay was determined using fish samples collected monthly by a surrounding net from October 2008 to September 2009. A total of 21 species, 582 individuals and 5,345.1 g of fishes were collected during the study period and Perciformes accounted for 47.6 % of the total number of species. Takifugu niphobles, Engraulis japonicus and Lateolabrax japonicus predominated in number, accounting for 55 % of the total number of individuals. Higher number of species and individuals occurred in June and in September-October. Species were grouped into three by cluster analysis: A) dominant species such as Leiognathus nuchalis and Takifugu niphobles, B) fish collected mainly in spring such as Sebastes inermis, Chaenogobius heptacanthus, Pterogobius zonoleucus, Pholis nebulosa, and Trachyrhamphus serratus, and C) fish collected mainly in autumn such as Engraulis japonicus, Lateolabrax japonicus, and Hyporhamphus sajori. 가막만 모자반 군락에서 2008년 10월부터 2009년 9월까지 월별로 두릿그물로 어류를 채집하여 어류의 종조성 및 계절변동을 분석하였다. 조사기간 동안 21종 582 개체, 5,345.1 g의 어류가 채집되었으며, 출현한 어종 중 복섬(Takifugu niphobles), 멸치(Engraulis japonicus)와 농어(Lateolabrax japonicus)가 전체 개체수의 55 %를 차지하였다. 12월과 1월에는 어류가 채집되지 않았고, 봄에 수온이 상승하며 증가하여 출현종수와 채집량이 6월에 높았고, 7월과 8월에는 감소하였으나, 9월과 10월에는 높은 값을 보였다. 종간 유사성을 집괴분석한 결과 3개의 무리로 구분되었다. A 그룹에는 출현기간이 비교적 긴 우점종인 주둥치(Leiognathus nuchalis)와 복섬(Takifugu niphobles)이, B 그룹에는 봄에 주로 출현한 볼락(Sebastes inermis), 살망둑(Chaenogobius heptacanthus), 흰줄망둑(Pterogobius zonoleucus), 베도라치(Pholis nebulosa), 거물가시치(Trachyrhamphus serratus), C 그룹에는 가을에 주로 출현한 멸치(Engraulis japonicus), 농어(Lateolabrax japonicus), 학공치(Hyporhamphus sajori)가 포함되었다.

Purification and Characterization of Alu I Methylase

윤호섭,서향,한문희,유욱준,Yoon, Ho-Sup,Suh, Hyang,Han, Moon-H.,Yoo, O.-Joon 생화학분자생물학회 1985 한국생화학회지 Vol.18 No.1

Alu I methylase를 순수 정제하였다. 300g (wet weight)의 Arthrobacter luteus에서 얻은 crude extract로부터 ammonium sulfate fractionation을 거친 후 phosphocellulose, DEAE-cellulose, Heparin-agarose, Hydroxylapatite등의 chromatography과정을 통하여 0.88mg의 Alu I methylase를 얻었다. Specific activity는 mg당 $1.32{\times}10^5\;unit$이었다. 이 methylase에 의하여 methylation된 DNA는 Alu I endonuclease에 의하여 절단되지 않았으며, 그 methylation site는 5'-d($AG^{\downarrow}CT$)-3'임이 확인되었다. 정제된 Alu I methylase는 10% SDS-polyacrylamide gel electrophoresis에서 한개의 major band로 나타났으며 그 분자량은 $56,000{\pm}1,000$ 이었다. Alu I methylase has been isolated from 300g (wet weight) cells of Arthrobacter luteus. After ammonium sulfate fractionation, the protein which has methylase activity was purified through phosphocellulose, DEAE-cellulose, Heparin agarose, and Hydroxy-lapatite column chromatography. The methylated DNA by the purified methylase was resistatnt against Alu I endonuclease. The purified Alu I methylase was essentially homogeneous as judged by 10% SDS-polyacrylamide gel electrophoresis, and the apparent subunit molecular weight was $56,000{\pm}1,000$. The specific activity of the enzyme was $1.32{\times}10^5$ units per mg protein.

Alu I Methylase 의 정제와 특성

윤호섭,서향,한문희,유욱준 ( Hosup Yoon,Hyang Suh,Moon H . Han,O . Joon Yoo ) 생화학분자생물학회 1985 BMB Reports Vol.18 No.1

Alu I methylase has been isolated from 300g (wet weight) cells of Arthrobacter luteus. After ammonium sulfate fractionation, the protein which has methylase activity was purified through phosphocellulose, DEAF-cellulose, Heparin agarose, and Hydroxylapatite column chromatography. The methylated DNA by the purified methylase was resistatnt against Alu I endonuclease. The purified Alu I methylase was essentially homogeneous as judged by 10% SDS-polyacrylamide gel electrophoresis, and the apparent subunit molecular weight was 56,000±1,000. The specific activity of the enzyme was 1.32×105 units per ㎎ protein.

눈 영역 추출에 의한 얼굴 기울기 교정

윤호섭,왕민,민병우,Yoon, Ho-Sub,Wang, Min,Min, Byung-Woo 대한전자공학회 1996 전자공학회논문지-B Vol.b33 No.12

This paper describes facial component detection and skew correction algorithm for face recognition. We use a priori knowledge and models about isolated regions to detect eye location from the face image captured in natural office environments. The relations between human face components are represented by several rules. We adopt an edge detection algorithm using sobel mask and 8-connected labelling algorith using array pointers. A labeled image has many isolated components. initially, the eye size rules are used. Eye size rules are not affected much by irregular input image conditions. Eye size rules size, and limited in the ratio between gorizontal and vertical sizes. By the eye size rule, 2 ~ 16 candidate eye components can be detected. Next, candidate eye parirs are verified by the information of location and shape, and one eye pair location is decided using face models about eye and eyebrow. Once we extract eye regions, we connect the center points of the two eyes and calculate the angle between them. Then we rotate the face to compensate for the angle so that the two eyes on a horizontal line. We tested 120 input images form 40 people, and achieved 91.7% success rate using eye size rules and face model. The main reasons of the 8.3% failure are due to components adjacent to eyes such as eyebrows. To detect facial components from the failed images, we are developing a mouth region processing module.

살조개, Protothaca jedoensis 자원조성을 위한 양식생물학적 연구 I. 난발생과 유생사육

윤호섭,김정,최상덕,Yoon, Ho-Seop,Kim, Jung,Choi, Sang-Duk 한국양식학회 2005 韓國養殖學會誌 Vol.18 No.4

살조개의 안정적인 인공종묘생산을 위한 기초 연구로서 수온별 난발생, 먹이생물, 밀도에 따른 유생의 성장과 생존율을 조사하였다. 살조개의 발생가능 수은은 $15{\~}30^{\circ}C$인 것으로 나타났으며 초기 D형 유생에 이른시간은 $15^{\circ}C$에서 $30^{\circ}C$까지 각각 39.7, 31.2, 26.8, 26.2시간이 소요되었다 수온별 유생사육에서는 $30^{\circ}C$에서 성장과 생존율이 가장 양호하였다. 사육밀도에 따른 성장과 생존율은 4${\~}$6 ind./ml의 밀도에서 가장 적합한 것으로 나타났다. 식물먹이생물에 따른 성장과 생존율은 Isochrysis galbana, Pavlova lutheri, Chaetoceros calcitrans를 혼합 공급하는 것이 가장 효과적이었다. 먹이생물농도에 따른 성장과 생존율은 $1{\times}10^4$ cells/ml에서 $218{\mu}m$와 $45\%$로 가장 높았다. In order to obtain the aquaculture fundamental data far resources enhancelnent of the Protothaca jedoensis, the egg development and larva growth were investigated at different conditions such as water temperature, phytoplankton and density. Water temperature, at which P. jedoensis egg successfully completed development, ranged from $15{\~}30^{\circ}C$. The required time from fertilization to D-shaped larva was 39.7 hours at $15^{\circ}C$, 31.2 hours at $20^{\circ}C$, 26.8 hours at $25^{\circ}C$ and 26.2 hours at $30^{\circ}C$ P. jedoensis. In regard to water temporature, growth and survival rates of larvae were high at $30^{\circ}C$. In growth and survival rates of larvae with various rearing densities, the highest aver-age growth and survival rates were 4${\~}$6 ind./ml When larvae were fed mixed phytoplankton, such as Isochrysis galbana, Pavlova lutheri and Chaetoceros calcitrans, their growth and survival rates were the high among the groups. In growth and survival rates of larvae with various rearing food concentrations, the highest average growth and survival rates were $218{\mu}m$, and $45\%$ at the food concentration of $1{\times}10^4$ cells/ml, respectively.

Alu I Methylase 의 효소적 특성과 작용부위

윤호섭,서향,김기태,한문희,유욱준 ( Hosup Yoon,Hyang suh,Ki tae Kim,Moon H . Han,O . Joon Yoon ) 생화학분자생물학회 1985 BMB Reports Vol.18 No.1

The specific methylation site for Alu I methylase was the cytosine nucleotide in Alu I sequence. The position of the methylated cytosine nucleotide was determined by the chemical cleavage reactions of the Maxam-Gilbert DNA sequencing procedure. As expected, the methylated cytosine nucleotide bands were disappeared on C+T and C lanes on 1296 sequencing gels. Alu I methylase was maximally active at near pH 7.5 in the presence of 50 mM NaCI. The methylase did not require Mg^(++) for activity, and obeyed Michaelis-Menten Kinetics with respect to both AdoMet and DNA. At 37℃, the Km for AdoMet was 0.44 μM, that for the Alu I site of pBR 322 DNA was 4.03 nM, and the corresponding turnover numbers were 1.83 methyl transfer per minute per monomer and 1.61 transfers per minute per monomer, respectively.

The Specificity and Catalytic Properties of Alu I Methylase

윤호섭,서향,김기태,한문희,유욱준,Yoon, Ho-Sup,Suh, Hyang,Kim, Ki-Tae,Han, Moon-H.,Yoo, O.-Joon 생화학분자생물학회 1985 한국생화학회지 Vol.18 No.1

순수 정제된 Alu I methylase (Yoon et al., 1985)의 specificity와 그 효소적 특성을 연구하였다. 이 효소는 DNA의 염기 배열 5'-d($AG{\downarrow}CT$)-3'을 인식 하였으며 그 중 cytosine nucleotide에 methylation을 시키고 있음이 Maxam-Gilbert의 방업에 의한 DNA sequencing을 통하여 밝혀졌다. Alu I methylase는 Alu I site 당 두 개 의 methyl group을 붙여 주였으며 Eco RI에서와 같은 overmethylation현상은 보여주지 않았다. 효소반응은 pH 7.4에서 7.6사이, NaCl은 50 mM에서 최적 이었다. Alu I methylase는 Michaelis-Menten kinetics를 따랐으며 S-Adenosyl-methionine에 대한 $K_m$ 값은 $0.44\;{\mu}m$이었고 pBR 322 DNA를 사용 하였을 때의 DNA에 대한 $K_m$의 값은 $4.03\;{\mu}m$이었다. 이 $K_m$값들로부터 얻어진 turnover number는 각각 1.83/min과 1.61/min이었다. The specific methylation site for Alu I methylase was the cytosine nucleotide in Alu I sequence. The position of the methylated cytosine nucleotide was determined by the chemical cleavage reactions of the Maxam-Gilbert DNA sequencing procedure. As expected, the methylated cytosine nucleotide bands were disappeared on C+T and C lanes on 12% sequencing gels. Alu I methylase was maximally active at near pH 7.5 in the presence of 50 mM NaCl. The methylase did not require $Mg^{++}$ for activity, and obeyed Michaelis-Menten Kinetics with respect to both AdoMet and DNA. At $37^{\circ}C$, the $K_m$ for AdoMet was $0.44\;{\mu}m$, that for the Alu I site of pBR 322 DNA was 4.03 nM, and the corresponding turnover numbers were 1.83 methyl transfer per minute per monomer and 1.61 transfers per minute per monomer, respectively.

시간 에지와 컬러 정보를 이용한 손 영역 자동 추적

윤호섭(Ho-Sub Yoon),소정(Jung Soh),왕민(Min Wang),민병우(Byung-Woo Min) 대한전자공학회 1997 대한전자공학회 학술대회 Vol.1997 No.6

보강재(whisker)를 이용한 플라스틱 복합재료의 특성 및 초미세 발포를 이용한 수축률 제어

윤호섭(H.S. Yoon),손주성(J.S. Sohn),유영재(Y.J. Ryu),차성운(S.W. Cha) 한국정밀공학회 2018 한국정밀공학회 학술발표대회 논문집 Vol.2018 No.5월