백서 뇌에서 전기경련 충격에 의해 발현이 조절되는 새로운 유전자

정선주,박정은,강웅구,구영진,박주배,김용식 大韓神經精神醫學會 2000 신경정신의학 Vol.39 No.5

연구목적: 이 연구는 백서 뇌에서 전기경련 충격(Electroconvulsive shock, ECS)에 의해 발현이 조절되는 새로운 유전자를 검색하고, 발달단게 및 뇌 조직에 따른 발현양상을 관찰하기 위하여 시행되었다. 방법: 생후 7일, 21일된 어린 웅성 백서에 전기경련 충격을 가한 후 mRNS differential display-PCR(DD-PCR) 기법을 시행하여 발현의 차이를 나타내는 유전자를 검색한 후 서열분석을 실시하였다. 생후 7일 및 21일된 백서의 뇌와 성년 백서의 대뇌피질, 해마를 이용한 Northern blot 분석을 시행하여 검색된 유전자의 ECS 처치 후 시간경과에 따른 유전자 발현양상을 관찰하였다. 결과: ECS 처치군과 위충격군간에 발현의 차이를 나타낸 유전자 4개를 검색할 수 있었으며, 이중 ECS에 의해 발현이 증가되는 1개의 유전자를 선정하여 서열분석을 실시한 결과 EST 서열만이 보고된 새로운 유전자임이 밝혀졌다. 이 유전자는 생후 7일된 백서의 뇌에서는 ECS 처치 후 180분에 발현양의 증가가 최고치에 도달하였으나, 생후 21일된 백서의 뇌와 성년백서의 대뇌피질 및 해마에서는 360분 경과시까지 지속적으로 발현 양이 증가하였다. 결론: 이같은 결과는 ECS에 의해 생체 내에서 유전자 발현의 조절이 이루어짐을 시사하며, 향후 이러한 유전자의 세포 내 기능에 관한 연구를 시행함으로써, ECS에 의한 생체의 반응과 치료기전에 대한 이해를 넓힐 수 있을 것으로 기대된다. 중심단어:전기경련 충격·DD-PCR·유전자·백서 뇌. Objectives: This study was performed to identify genes regulated by electroconvulsive shock (ECS) and to observe the pattern of expression of genes according to different developmental stages and brain regions. Method: ECS(130V, 0.5 sec) was given to male Sprague-Dawley rats with age of postnatal day 7 and 21(P7, P21 respectively). After screening genes regulated by ECS with mRNA diffe-rential display-PCR(DD-PCR), we selected one clone among them and observed the induction of this gene after ECS by time-dependent Northern blot analysis of rat brain of P7, P21 and adult rat cortex and hippocampus. Results: By DD-PCR method, we have identified four clones whose expression was regulated by ECS. Among them, one(CP 10-2) was proved to be a new gene by sequencing and BLAST search. Its expression was increased after ECS in P7, P21, and adult rat brain. The expression of CP 10-2 reached peak level at 180 minutes after ECS in P7 rat brain, but was further increased until 360 minutes after ECS in P21 and adult rat brain. Conclusion: In this study, a new gene was identified in rat brain which showed up-regulated expression in response to ECS. Cloning and characterization of this new gene would be helpful to elucidate the effect of ECS in rat brain. KEY WORDS:Electroconvulsive shock·Differential display-PCR·Gene·Rat brain.

Vitreoscilla Catalase의 분리 및 특성에 관한 연구

박기인,박충웅 全北大學校 基礎科學硏究所 1994 基礎科學 Vol.17 No.-

Vitreoscilla는 헤모글로빈을 갖고 있는 그람 음성 미생물체로써, 이 헤모글로빈을 통한 산소호흡으로부터 파생된 과산화수소를 많은 양의 catalase를 합성하여 효과적으로 분해한다. 이 효소의 분자량은 약 250,000 daltons이며, subunit의 구조는 모든 다른 미생물체에서 보여주는 A_4와는 달리 A_2B_2(A:64,000 daltons, B:58,000 daltons)를 보였다. 최적 pH는 7.0∼8.0이었고, oxidized form의 absorption spectra에서 Soret peak는 406㎚에서 나타났으며 reducing agent인 sodium dithionite로부터 야기된 reduced form의 Soret peak는 442㎚로 이동되었다. 또한 이 효소는 열에 불안정 했고, K_m값은 0.016M hydrogen peroxide를 나타냈으며, tumover number는 25,000mol이었다. 0.25mM potassium cyanide는 경쟁적 저해로써 이 효소의 활성을 저해하였으며, 효소-저해제 복합체의 dissociation constant는 0.67mM을 보였다. 두 subunits의 N-말단 아미노산 서열이 다음 실험을 위한 oligonucleotide 합성을 위하여 결정되었다. Vitreoscilla is a gram-negative bacterium that contains a unique bacterial hemoglobin and grows very well under the condition of low oxygen concentration. It also contain a bacterial catalase to be not correspond with another species on genus Beggiatoa. The primary function of Vitreoscilla catalase may be to remove hydrogen peroxide produced by VitHb oxidation. The molecular size of the catalase was estimated to be approximately 250,000 Da. The subunit structure of this enzyme may be A_2B_2 (A: MW 64,000 Da, B: MW 58,000 Da) but is not clear in the research reported here. Optimum pH is 7.0∼8.0 for catalase activity and Soret peak on absorption spectra of oxidized catalase is represented in 406㎚ and Soret peak of reduced form from sodium dithionite moved at 442㎚. Vitreoscilla catalase is unstable a high temperature, and its Michealis constant, K_m, was 0.016 M hydrogen peroxide. The tumover number of the enzyme was 25,000mol. The 0.25mM potassium cyanide was competitive inhibitor and the dissociation constant of the enzyme-inhibitor complex was 0.67mM. N-terminal amino acid sequences of two subunits are determined for the probe synthesis using to the cloning of Vitreoscilla catalase gene.

Saccharomyces sp. 균주들을 이용한 감즙으로부터 알콜 생성에 대한 연구

朴忠雄,李康民,金景淑 全北大學校 1993 論文集 Vol.36 No.-

By using the Korea persimmon juice as raw material, persimmon wine fermentation was carried out with 5 strain of Saccharomyces (S. cerevisiae, S. formosensis, S. sake, S. calsberg, S.ellipsoideus). S.cerevisiae and S. formosensis are very suitable for conversion of persimmon juice to persimmon alcohol but not S. sake, S. calsberg, S. ellipsoideus. In the process of fermentation, the compositions of amino acids were changed. Aromatic amino acids such as phenylalanine and tyrosine were decreased but arginine and methionine were increased. Amounts of sugars(glucose, fructose) and Saccharomyces affect on persimmon alcohol concentration produced. Persimmon alcohol concentration was double with adding of 10% glucose in S. formosensis fermentation. In S. cerevisiae fermentation, glucose could not augment the alcohol concentration. Finally we got 11.5% persimmon alcohol by S. formosensis. We designed a alcohol bioreactor by immobilization of S. formosensis on porous supporter and packing it in column.

Chlorella Protein의 營養評價에 關한 硏究〔Ⅱ〕 : 人工消化率과의 比較 Comparative of Artificial Digestion Ratio

朴忠雄,黃鎬觀 全北大學校 1974 論文集 Vol.16 No.-

The excellency of nutritional value in Chlorella Protein had already been proved, in the previous report of mine, by animal metabolic test. This time, by artificial digestive test, the following results have been obtained : 1) The artificial digestive ratio is increased by 70%, when 1(one) mg of Pepsin is acted on 1(one) gram of sample. 2) The digestive ratio is increased by 10% when the sample is homogenized. From this, we could see that the digestive ratio for Chlorella Protein is dependent upon the previous processes. 3) The artificial digestive test after the process of homogenation has shown us that Chlorella, when cultured by the synchronous culture method, is good in its quality, though its harvest per unit area is small in quantity.

工業專門大學 食品工業科 敎育課程 開發에 關한 硏究

朴正吉,柳寅德,李雄洙,趙載敏 忠州大學校 1982 한국교통대학교 논문집 Vol.15 No.2

This is a study on the improvement of the curriculum of food technology department in technical junior college in parallel with the development of scientific technique. The results obtained is as follows; 1. It is emphasised with training the middle-level technician in both quality control and food processing course in curriculum. 2. It is established with new subject; Instrumental Analysis, EDPS, Introduction to Electric Technology, Industrial Management, Quality control. 3. With elimination of overlapping subjects it is assembled with 18, 19 subjects in each course. 4. It is suggested with the topic of experiments and practices extracted from the result of job analysis.

原子吸光法에 依한 畓土壞의 미량원소 定量

朴忠雄 全北大學校 文理科大學 1977 論文集 Vol.5 No.-

This experiment was discussed in term of varing the absorption of Atomic Absorption Spectrophotometry due to the analysis of initial sample, the effect of pH, and interference of other elements. It was reported that the absorbance of potassium was increased and the absorbance of calcium, magnesium and zinc was decreased because of interference of Cu^++ and Fe^++, that the absorbance of potassium, calcium, magnesium and zinc was varied with effect of pH. The results obtained in this experiment are summarized as follows : 1. Optimal condition of pH in paddy soils was neutral state. 2. The absorbance of potassium was increased and the absorbance of calcium, magnesium and zinc was decreased by adding of Fe^++ and Cu^++ in the initial sample solution.

새로운 LacZ Gene Fusion 및 Gene Cartridge Vector의 개발에 관한 연구

박교선,이영훈,박충웅,최창진 圓光大學校 基礎自然科學硏究所 1987 基礎科學硏究誌 Vol.6 No.2

In this study, a new lacZ gene vector, plasmid PGEM3-lacZ was constucted, which could be used for the study of the gene regulation and the function of proteins. Plasmid pMC1871 was digested with restriction endonuclease Pst I and Sma I to obtain a DNA fragment with the lacZ-gene(3091 bp). Recombinant plasmid pGEM3-lacZ was constructed by inserting the lacZ gene DNA(Pst I-Sma I cut DNA; 3091 bp) of pMC1871 into the Pst I and Hinc Ⅱ sites of plasmid pGEM3. After then, Recombinant plasmid pGEM3-lacZ DNA was transformated into E. coli HB 101. pGEM3-lacZ DNA/HB 101 was screened on the NZY plate with ampicillin and X-gal(5-Bromo-4-Chloro-3-Indolyl-β-D-Galactoside) by selecting blue colonies. The restriction map of pGEM.3-1acZ DNA was constructed by the following restriction enzymes :Bgl I, Xbal I, Bam HI, EcoR I, Hind Ⅲ, Kpn I, Pst I, Sma Ⅰ, Sal Ⅰ. From the restriction map, the lacZ DNA was confirmed to be present at the expected position in the plasmid pGEM3-lacZ. pGEM3-lacZ/HB101 gives about 10 times more lacZ DNA than pMC 1871/HB101.

산업대학 건축공학과 교육과정 개발에 관한 연구

임응찬,한인웅,이승준,박재호,조방현,서형수,정구용,한기원,김진원,임상규,이재윤 三陟大學校 1996 論文集 Vol.29 No.2

By impproving the educational program of the Department of Architectural Engineering and studying the theory of learning and the applied method necessary for the development of nation and society, we should develop a satisfying course of education to meet the demand and skill in the industralized society for the purpose of cultivating men of baility with great personalities and creative minds. Therefore, this study focused on classifying several kinds of occupations in the prart of architectured related to what graduates from the department of Architectural Engineering go with into the society. Accordingly after choosing some works adequate to each occupation by the proposed curricula, we tried to establish the scope of education and courses corresponding to the jobs and thus and develop a satisfying curriculum.

海水를 含有한 廢水의 微生物 轉煥에 關한 硏究

韓雄傳,朴淸吉 釜山水産大學校 1980 釜山水産大學 硏究報告 Vol.20 No.2

Effects of chloride on the biological treatment process of wastewater containing seawater was studied. The dextrose-synthesized wastewater and the synthesized waste-waters containing different chloride concentrations of 2,760, 8,460 and 16,690 ppm, which were adjusted by adding seawater to the dextrose-synthesized wastewater, were treated, respectively, by the aeration-only complete mixing activated sludge(CMAS) process to evaluate the chloride effects on the values of the kinetic constants, organic substrate removal rate and the effluent quality. The fish-processing wastewater whose T-BOD and chloride concentration were 1,551 and 4,450㎎/ℓrespectively was also treated by the complete mixing activated sludge process with return sludge to estimate the kinetic constant values, organic substrate removal rate and effluent quality. The units of the aeration-only CMAS process were operated with the hydraulic detention time variations of from 0.133 to 0.58 day while that of CMAS process with return sludge was operated with the hydraulic detention time variations of from 0.18 to 9.42 day and cell residence time variations of from 1.10 to 2.5 day. The kinetic constant values of k, Y, ??, for the synthesized wastewater were estimated to be ??, 0.69 and ?? respectively. These values were not changed by the increases in chloride concentration in the synthesized wastewater. However, the Ks value only varied from 83.3 to 139.5㎎/ℓas the chloride concentrations in the wastewater increased. The kinetic constant values of Ks, k, Y and ?? for the fish-processing wastewater were found to be 257㎎/ℓ, ??, 0.54 and ??, respectively. And the oxygen requirement per biomass for endogeneous respiration(b´) were estimated to be 0.25 and ?? respectively. The value of BOD coefficient(Kb) in the waste-water containing seawater were 0.72, which is slightly lower than that of the synthesized wastewater. It was suggested that design parameters for treating the high chloride ion-containing wastewater be evaluated based on the MLVSS rather than MLSS from the fact that the ratio of MLVSS and MLVSS and MLSS was only 0.49. In treating the fish-processing wastewater by CMAS process with MLVSS concentration of 4,000 ㎎/ℓ, it was found that the hydraulic detention time and cell residence time should be greater than 6.2 hours and 1.5 days to meet the effluent quality, respectively.

Dog Liver Mevalonate Kinase의 分離및 成質에 關한 硏究

趙仁鎬,朴忠雄 全北大學校 基礎科學硏究所 1981 基礎科學 Vol.4 No.1

Mevalonate kinase(EC 2. 7. 1. 36), the first enzyme in the ATP-requiring sequence of cholesterol bisoynthesis, has been purified from dog liver and investigated some its properties. 1. This enzyme has a specific activity of 27. 21 μmoles of product formed per min per mg of protein. 2. The effect of MG^2^++, Mn^++ on MVA kinase activity was investigated as follw; Mn^++> Mg^++(at low concentration) Mg^++> Mn^++(at high concentration) 3. The initial velocity pattern suggests that the mechanism of the dog liver mevalonate kinase reaction sequential.