兩棲類 三種의 同位酵素와 核型에 관한 硏究

鮮于 洋一,嚴慶一 東亞大學校 大學院 1981 大學院論文集 Vol.5 No.-

Isozymes and chromosomes of Kaloula borealis, hyla alborea japonica, and Bombina arientalis were studied by starch gel electrophoresis and bone marrow air-drying method. Twenty-three enzymatic proteins and enzymes of the three anuran species collected in Seoul province provided a basis for estimating the genetic distance among these species. On the genetic similariry, the relationship between Bombina orientalis and Kaloula alborea was nearer than the others. The chromosome number of Hyla alborea japonica and Bombina orientalis was 24, and Kaloula borealis was 28. The differences of these were a little.

균체외단백질 생산균 Bacillus brevis P176-2 균주의 동정과 배양조건

채기수,엄경일 東亞大學校大學院 1991 大學院論文集 Vol.16 No.-

Identification and influence of the cultural conditions upon extracellular protein production by Bacillus brevis P176-2 isolated from soil were investigated. The morphological, physiological and cultural characteristics of strain P176-2 were studied according to the methods of Bergey's manual. Based on the results obtained in thses experiments, strain P176-2 was identified as Bacillus brevis. The extracellular protein ws efficiently produced when added to 1.0% peptone as nitrogen source. Optimum initial pH and temperature for extracellular protein production were 7.5 and 40℃ respectively. Higher concentration of magnesium ion stimulated growth, but prevented extracellular protein production. Optimum concentration of potassium phosphate was found to be 0.2%. This strain produced 5.38mg/ml of extracellular protein when cultured for 48 hours under optimum conditions. The extracellular protein produced under optimum conditions contained relatively large amounts of aspartic acid, lysine and arginine, but cystine and methionine were very low.

Environmental Toxic Agents on Genetic Material and Cellular Activity : Ⅲ. 포유동물세포에서 돌연변이원에 의한 DNA상해의 회복에 미치는 DNA 중합효소저해제의 영향 Ⅲ. DNA Polymerase Inhibitors on Repair of Mutagen-Induced DNA Damage in Mammalian Cells

李天馥,嚴慶一,鮮于洋一,申恩珠 東亞大學校附設基礎科學硏究所 1988 基礎科學硏究論文集 Vol.5 No.1

Many enzymes are known to be associated with the recognition of specific lesions and incision of the DNA strand in their vicinity(Clarson and Mitchell, 1983), and several enzymes concerned with excision and polymerization have also been identified(Collins & Johnson, 1984; Collins et al., 1984; Downes et al., 1985; Mattern et al., 1982). In a number of studies, controversial results have been obtained on whether DNA polymerase α is(Cleaver, 1984; Collins et al., 1982; Dresler and Frattini, 1986: Dresler and Kimbro, 1987; Johnson et al., 1987; Snyder and Reagan, 1981, 1982) or is not(Giulotto and Mondello, 1981; Hardt et al., 1981; Seki et al., 1980) important in DNA repair, while DNA polymerase β has been postulated to have a role in the repair of DNA(Cleaver, 1983; Dresler and Lieberman, 1983; Miller and Chinault, 1982; Spadari et al., 1982). Many conclusions concerning the functions at these enzymes can be obtained using enzyme inhibitors. Among them, the commonly used inhibitors have been aphidicolin(APC), an inhibitor of DNA polymerase α(Smith and Paterson, 1983; Th'ng and Walker, 1985), and 2',3'-dideoxythymidine(ddThd) or 2',2'-dideoxythymidine 5'-triphosphate(ddTTP), an inhibitor of DNA polymerase β(Dresler and Kimbro, 1987; Th'ng and walker, 1985; Yamada et al., 1985). On the other hands, the studies using APC and ddTTP yielded results that suggest the participation of both polymerases in DNA repair depending on the agent used and the dosage applied(Cleaver, 1983; Dresler and Liberman, 1983; Yamada et al., 1985). Therefore, the same approach, using three differenc assays for DNA repair and two inhibitors, has been undertaken in this study with the view to give insight into the role of specific enzymes in the repair process. The purpose of this study is to elucidate the involvement of DNA polymerase α or β in DNA repair synthesis induced by EMS, an alkylating agent, or BLM, an antibiotics and X-ray mimetic agent, in CHO cells.

전기천공법에 의한 Bacillus brevis P176-2의 형질전환

채기수,엄경일 東亞大學校附設基礎科學硏究所 1993 基礎科學硏究論文集 Vol.10 No.1

균체외단백질 생성균 Bacillus brevis P176-2 균주를 pUB110 plasmid DNA로 전기천공법에 의한 형질전환을 시도하여 본 방법의 유효성과 최적조건을 검토하였다. 본 실험에 사용한 전기천공 장치로 전장강도 12.5kV/㎝, capacitor 용량 7.3㎌의 조건하에서 고전압 pulse를 세포에 가했을 때 가장 높은 형질전환율(1.26× 10⁴/㎍ plasmid DNA)을 나타내었으며 세포 생존율은 10% 정도이었다. 세포의 생육도가 ??에서 약 1.2정도로 증식하였을 때 세포를 수확하여 형질전환 시키는 것이 가장 좋았으며 그 이상의 생육도에서는 세포의 생존율은 증가되었지만 형질전환율은 낮아졌다. 전강강도 12.5kV/㎝, capacitor용량 7.3㎌ 조건의 pulse를 세포에 2회 연속적으로 가할 때 최대의 형절전환율(2.40× 10⁴/㎍ plasmid DNA)을 보였다. 세포의 생존과 pulus의 지속시간에 영향을 주는 electroporation burrer는 7 mM HEPES(pH 7.4), 272mM sucrose 및 1mM MgCl₂을 함유하는 buffer를 사용하여 세포를 수확하고 현탁하여 형질전환할 때 형질전환율과 세포 생존율이 가장 높았다. 세포현탁액에 첨가되는 plasmid DNA의 농도가 10㎍/ml일 때 까지는 농도에 비례하여 형질전환율이 높아졌으나 그 상의 농도에서는 비례하지 않았다. The optimum conditions and mechanisms for the plasmid-mediated genetic transformation of intact cells of bacillus brevis P176-2, an extracellular protein producing bacterium, by electroporation were investigated. It was found that pUB110 plasmid DNA can be introduced into intact bacterial cells by electroporation. The frequency of transformation by this electroporation system depended upon the initial electric field strength, the capacity of the clectric discharge capacitor, growth stage, number of successive pulses and composition of electroporation buffer. It was effective for transformation that cells were harvested, washed and resuspended with HSM[7mM HEPES(pH 7.4), 272mM sucrose, 1mM MgCl₂] electroporation buffer when cell growth was attained to 1.2 at ?? A maximum frequency of transformation of 2.40× 10⁴ transformants per ㎍ plasmid DNA was obtained by two succesive pulses with an initial electric field strength of 12.5kV/㎝ and with a capacitance of 7.3㎌. Key words : Bacillus breves, pUB110, plamid DNA, transformation, electroporation.

Chinese hamster 난소세포에서 DNA - 단백질 교차결합과 glutathione S - 전이효소의 발현에 대한 돌연변이원에 의해 유도된 교차적응반응

이동욱,엄경일,오규선,장정현,문용석 한국유전학회 2001 Genes & Genomics Vol.23 No.2

The present study has been undertaken to elucidate whether glutathione S-transferase (GST) is related to cross-adaptive response to DNA-protein crosslink (DPC) induced by ethyl methanesulfonate (EMS) or ultraviolet radiation (UV)-C in Chinese hamster ovary (CHO) cells. DPC was determined by SDS-K^+ precipitation method, and GST was measured by western blot analysis. 2,4-Dinitrophenol (DNP) that depletes cellular ATP and cycloheximide (CHM) that inhibits protein synthesis were used to find whether DNP and CHM affected the expression level of GST and cross-adaptive response to EMS or UV. There is a cross-adaptive response to DPC in cells pretreated with EMS or UV and subsequently treated with UV or EMS, respectively. In cells pretreated with 2 mM EMS and subsequently treated with 5J/㎡ UV, DNP and CHM did not affect cross-adaptive response to DPC and GST. And also, in cells pretreated with 1J/㎡ UV and subsequently treated with 8 mM EMS, CHM did not affect crass-adaptive response to DPC and the expression level of GST. But DNP increased the amount of DPC, and decreased the expression level of GST in UV-pretreated cells. These results suggest that GST are involved in the induction of cross-adaptive response to DPC in CHO cells.

경찰공무원의 건강체력 실태

노성규,손호중,고성은,엄경일,권호준 江原大學校附設體育科學硏究所 2003 江原大學校附設體育科學硏究所論文集 Vol.- No.26

The purpose of this study is to suggest basic data to make out the program about the health and promotion of physical fitness. So, we measured the status of the present police officers physical fitness through measuring their height, weight, %body fat, and healthy physical fitness and through comparing it with popular physical fitness survey data in an age. The conclusion that compared and analyzed the measuring result of the height, weight, and the element of healthy physical fitness of 143 police officers in G province is following First, Their height, weight, board jumping and VO2max is seemed to be high as compared with the standard. Second, %Body fat is closely similar with the standard. Third, The flexibility and sit-up is seemed to standard below By this time, it is true that the study of the superiority players evaluation of physical fitness is developing in our country. But, the study of the physical fitness evaluation about non-player, that is general person has been neglected in korea. So, it is not very enough the study and developing program for the improving the general person's physical fitness. Therefore, It should make the spacial study of the various physical fitness of korean in an occupation, a sex and age. And then, the special exercise program that can satisfy the various needs of today 'men is needed. Also, it should make the vertical study for the improvement of physical strength in vocational classification. Also, the government and office should invest institutional effort to help the police officer' health and welfare. Police officers also have to learn themselves the efficient and continuous exercise habit to be happy with health in considering their own physical condition. And then, the prescription of the exercise program about the result must come with.

Reduced Addamycin Cytotoxicity in RIF-1 Multicell Spheroid Due to an Acidic Microenvironment

Um, Kyung-Il,Cheston, Sally B.,Suntharalingam, Mohan,Rhee, Juong-G. Korean Environmental Mutagen Society 1997 한국환경성돌연변이·발암원학회지 Vol.17 No.1

Variations in adriamycin uptake and cytotoxicity were studied in tumor cells that were grown in different growth states and microenvironments. RIF-1 tumor cells were maintained in an RPMI 1640 medium, and grown in either a monolayer or multicell spheroids. For exponentially growing cells, adriamycin cytotoxicity increased with increased dosage up to 2.5 $\mu$g/ml, and this cytotoxicity was reduced when the cells were grown in a plateau phase or in an acidic microenvironment (pH 6.6). This reduced cytotoxicity was correlated with the uptake of the drug. For multicell spheroids, the cytotoxicity of the drug was reduced dramatically, and this reduction was also correlated with a reduced uptake of the drug and an acidic pH inside of the spheroids. When the drug cytotoxicity was evaluated at different locations within the spheroids, the cells in the inner regions were least affected by the drug, suggesting that both an acidic microenvironment and noncycling plateau phase cells are contributing factors in decreasing the efficacy of the drug in an organized tissue, such as multicell spheroids.

Effects of Environmental Toxic Agents on Genetic Material and Cellular Activity : Ⅱ. S-phase Dependent Sister Chromatid Exchanges and Chromosome Aberrations Ⅱ. DNA合成시기에서의 염색체 이상과 자매염색분체 교환

Um,Kyung Il,Choe,Jin Hong,Kim,Choon Kwang 東亞大學校附設基礎科學硏究所 1988 基礎科學硏究論文集 Vol.5 No.1

Evidence has been presented that sister chromatid exchanges(SCEs) of mitotic chromosomes are formed exclusively during the period of DNA synthesis(Kato, 1974; Wolff et al., 1974). DNA is synthesized after a special initiating event at a replicating origin creats a DNA replication fork. Replication fork origins tend to be activated in clusters called replication units or replication clusters(Hand, 1978). Painter(1980) suggested that the unit of SCE formation is identical with the cluster. It has also known that the time of replication and the size of replication clusters are regulated by a specific program(Hand, 1978). Therefore, the induction of SCE can be studied by modification in the schedule of the replication processes using various agents that inhibit DNA synthesis(Rainaldi et al., 1984). The morphology of prematurely condensed chromosomes(PCC) is various according to the stage of the interphase cells in the cell cycle at the time of fusion: The PCC of G1-phase are very long single chromatids, those of G2 are clongated and slender double chromatids and those of S are characterized by pulverized appearances. Those facts indicate that the induction of PCC promises to be useful for a number of purpose that are not directly related to the study of mitosis(Johnson and Rao, 1970; Ringertz and Savage, 1976). Mitomycin-C(MMC) is group of anti-tumor, antibiotics produced by Streptomyces caespitosis(Hata, 1956). It is known a cross-linkin agent (Natarajan and Obc, 1978) and is efficient in inducing SCEs(Gebhart, 1982). Bleomycin(BLM) is a group of water-soluble, basic glycopeptides and collected from Streptomyces verticillus(Umezawa et al., 1966). It is known to act like X-rays that induce large numbers of chromosome aberrations but few SCEs(Hittelman and Rao, 1974; Gebhart, 1982). 1-β-D-arabinofuranosylcytosine(ara-C_ is a nucleside analog of 2-deoxycytosine in which the deoxyribose is replaced by arabinose and its predominant effect is a strong inhibitor of DNA synthesis(Benedict and Johnson, 1979). The purpose of this study was to investigate the induction of SCEs and its relation with chromosome aberrations induced by MMC and BLM using Ara-C in S-phase synchronized Chinese hamster ovary(CHO) cells.

Excision Repair Pathway by UV-light and Methyl Methanesulfonate in Chinese Hamster Ovary Cells

Um, Kyung-il,Sunwoo, Yang-il 東亞大學校 大學院 1984 大學院論文集 Vol.9 No.-

본 연구는 자외선과 Methyl methanesulfonate(MMS)를 복합 처리하여 이들에 의한 DNA 상해와 회복 경로를 규명하고, 아울러 이들에 의한 절제 회복과 DNA 사절단에 어떻게 상호 작용하는 가를 규명할 목적으로 수행하였다. 실험 결과는 다음과 같다. 1. MMS와 자외선을 복합 처리할 경우 이들에 의한 DNA 단사 절단률은 단독 처리군의 합보다는 약간 낮으나, 각각의 절단률보다는 높아 Additive 효과를 보였다. 2. 자외선에 의해 상해된 DNA의 회복 합성률은 선량의 증가에 따라 30J/m2 까지 증가하였고, MMS에 의한 경우도 농도의 증가에 따라 2mM까지 증가하였으며, 그 후는 plateau를 이루었다. MMS와 자외선을 복합 처리한 경우 이들 양자에 의한 DNA 회복 합성률의 합보다 훨씬 낮을 뿐 아니라 자외선 단독군의 값보다 낮았으나 MMS 단독 처리군보다는 높았다. 이는 MMS가 자외선에 의해 상해된 DNA의 회복 합성률을 억제하나, 자외선은 MMS에 의해 유발된 DNA 회복 합성률을 억제하지 않기 때문인 것으로 생각된다. 3. 자외선에 의해 유발된 피리미딘 이량체는 시간이 경과함에 따라 절제되어 조사 후 48시간에는 조사 직후의 약 50%만이 남았다. MMS 처리 후 자외선을 조사한 경우는 피리미딘 이량체의 절제가 억제되어 자외선 단독 처리 구분보다 더 많이 남아 있는 것으로 나타났다. 이는 MMS가 자외선에 의한 DNA 손상의 회복 합성률을 억제하기 때문이라 생각된다. 이상의 결과는 자외선에 의해 유발된 절제 회복에 관여하는 단백질 복합체가 절제 및 재삽입 단계에서 MMS에 의해 불활성화 되어질 수 있음을 시사한다.

DNA Repair Synthesis Induced by Bleomycin in HeLa $S_3$ Cells Pretreated with Base Analogs

Um, Kyung-Il,Park, Sang-Dai The Korean Society for Integrative Biology 1977 동물학회지 Vol.20 No.1

Bleomycin에 의해 유발된 DNA 회복합성은 저농도 처리군에서는 농도의 증가에 따라 증가하며 $5\\mu$g/ml 군에서 조사한 전세포의 15%가 회복합성을 하여 최고율을 보인다. 고농도 처리군에서 DNA 회복합성율이 감소하며 처리 시간을 연장해도 그율은 변화가 없다. BUdR이나 IUdR을 전처리한군에서는 DNA회복합성을 증가시키는 것으로 판명됐으며 또한 고동도 처리군에서는 정상적인 DNA 합성을 억제한다. 시간 변화에 따른 실험에서는 처리한 bleomycin을 제거한후 24시간까지 DNA 회복합성이 계속됐다. 이들 결과는 bleomycin이 excision repair를 유발하는 효과적인 화학물질이 아니며, bleomycin에 의해 유발되는 DNA의 손상은 DNA 나선 절단뿐만 아니라 다른 형태의 DNA 손상도 유발함을 추측할수 있다. Dose response of DNA repair synthesis induced by bleomycin was dose-dependent in lower doses, and maximum rate of it at 5 $\\mu$g/ml represents about 15% of total cells analyzed. At higher doses DNA-repair synthesis was reduced and the rate of it remained unchanged even prolonged treatment. Pretreatment with BUdR or IUdR was found to enhance DNA repair synthesis and also to interfere with semiconservative DNA synthesis at higher doses. Time dependence study showed that DNA repair synthesis occurred as long as for 24 hours after removal of bleomycin. These results seem to suggest that bleomycin is not to be an effective chemical in inducing excision repair and that damages induced in DNA by this drug might include not only strand breaks but other types of DNA damage.