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조류인플루엔자 H5N1 바이러스 유전자의 신속 검출을 위한 초고속 다중 실시간 PCR법의 개발
김을환,이동우,한상훈,임윤규,윤병수,Kim, Eul-Hwan,Lee, Dong-Woo,Han, Sang-Hoon,Lim, Yoon-Kyu,Yoon, Byoung-Su 대한수의학회 2007 大韓獸醫學會誌 Vol.47 No.4
Cause of high lethality and dissemination to human being, new development of rapid method for the detection of highly pathogenic Avian Influenza Virus (AIV) is still necessary. For the detection of AIV subtype H5N1, typical pathogenic AIV, new method to confirm sub-typing of this virus is also needed. For the purpose of ultra-rapid detection and sub-typing of hemagglutinin and neuraminidase of AIV, this study was planned. As the results we could demonstrate an ultra-rapid multiplex real-time PCR (URMRT PCR) for the detection of AIV In this study, the URMRT PCR were optimized with synthesized AIV H5- and AIV Nl-specific DNA templates and GenSpector TMC, which is a semiconductor process technology based real-time PCR system with high frequencies of temperature monitoring. Under eight minutes, the amplifications of two AIV subtype-specific PCR products were successfully and independently detected by 30 cycled ultra-rapid PCR, including melting point analysis, from $1{\times}10^3$ copies of mixed template DNA. The URMRT PCR for the detection of AIV H5N 1 developed in this study could be expected to apply not only detections of different AIVs, but also various pathogens. It was also discussed that this kind of the fastest PCR based detection method could be improved by advance of related technology in near future.
김을환,양태홍,이영자,최한영 서울保健大學 1990 論文集 Vol.10 No.1
This study was performed to investigate the content of a few inorganic metals and the degree of water temperature pH, DO, BOD, COD, SS, TH, Cl^(-), SO_(4)^(-), total alkali and PO_(4)^(3)-P in Singal Lake. In general they are favorable as the water to see but compared with the several terms which are measured in 1976 the pollution of the water increased by five or six times.
김을환 서울보건대학 1995 서울보건대학 부설 한국보건과학연구소 논문집 Vol.2 No.1
On this study the quality of water in the San-Jung lake was analyzed. The collection of the samples was performed in June 1994 by the Hyroth sampler at the upper, middle, lower strata of 5points selected in the San-Jung lake. The sample analysis was carried out was on the spot and in the laboratory. The result of this study was as follows ; 1. The saturation degrees of DO in the sample of upper stratum were 95.3∼100.7%, middle stratum 38.5∼96.1%, lower stratum 31.1∼92.0% respectively, whereas at the points S_(1), S_(2), S_(5), where each of the depth of water were specially deep, the saturation degrees of DO at the lower stratum(the nutrition decomposition stratum) were 30.1~33.1%. 2. TSI(SD) was 48.58, which meant that the trophic state was mesotrophic. 3. TSI (SD) was 74.26, which meant that the trophic state was eutrophic. 4. TSI(T-P) was 59.77, which meant that the trophic state was eutrophic. 5. The water quality condition in the San-Jung lake, due to the revised standard index of Carlson, was at the state of eutrophic.
상치 Phenolase에 의한 p-Coumaric Acid의 Hydroxylation
김을환,김연호 서울保健大學 1997 論文集 Vol.17 No.1
A phenolase from the leaves of lettuce(Lactuca sativa L.) was purified and some properties of its enzymic hydroxylation were studied. The experimental results are summerized as follows. 1. The lettuce phenolase catalyzes the hydroxylation of p-coumaric acid to caffeic acid in the presence of ascorbate. 2. The enzyme shows catalytic activity as hydroxylase and catechol oxidase in a constant ratio throughout the three stages of purification. 3. Ammonium sulfate and sodium chloride promote the activity of the enzyme; its maximum activity was observed in the 0.5M concentration of both salts. 4. Ascorbate functions as a reductant in quantities equivalent to the caffeic acid produced by hydroxylation. 5. 2-arnino-4-hydroxy-6, 7-dimethy1-5, 6, 7, 8-tetrahydropteridine and NADH can take the place of ascorbate, but caffeic acid can‘t.
HEMA와 DHPMA의 Copolymer에 대한 Urea, 저급 알콜 용질의 분배효과에 관한 고찰
김을환 서울保健大學 1991 論文集 Vol.11 No.1
A series of crosslinked 2-hydroxyethyl mechacrylate (HEMA)-2,3-dihydroxypropyl methacrylate (DHPMA) copolymer membranes for partition effect experiments has been prepared. The crosslinker used was tetraethylene glycol dimechacrylate(TEGDMA). Membranes were prepared by polymerizing the monomers as a thin homogeneous film. Using these membranes partition coefficients(KD) for low alcohols and urea were measured. This KD data shows that KD of isopropanol and urea increases as the DHPMA`s content increases when compose the HEMA-DHPMA copolymer.
Quick Real-time PCR을 이용한 Avian Influenza Virus Subtype H5N1의 신속검출법
김을환,이동우,한상훈,권순환,윤병수,Kim, Eul-Hwan,Lee, Dong-Woo,Han, Sang-Hoon,Kwon, Soon-Hwan,Yoon, Byoung-Su 한국미생물학회 2007 미생물학회지 Vol.43 No.1
조류 인플루엔자바이러스(AIV) H5N1 아형을 Real-time PCR법을 이용하여 가장 빠르게 진단할 수 있는 방법을 개발하였다. 검색 대상의 염기서열은 AIV H5N1 아형의 hemagglutinin 유전자 중 가장 상동성이 높은 387 bp의 부위를 선택하였고, 실험의 안전을 위하여 인공합섬의 방법으로 제작하였다. Microchip을 기반으로 한Real-time PCR법을 사용하였으며, 총PCR 반응액의 양을 $1{\mu}l$로, PCR 과정 중 각 단계, 즉 해리, 접합, 신장의 시간을 각1초, 1초, 3초로 하여 총 실험시간을 단축하였다. 진단을 위한 실험과정에서 PCR 및 융점분식에 소요된 최단 시간은 12분28초였으며, 민감도측정에서 최소2.4개의 hemaggutinin 유전자를 기질로 하여 목적한 특이 189 bp의 PCR 산물을 증폭할 수 있었기에, 본 연구에서는 이런 초고속 PCR 실험방식을 Quick Real-time PCR이라 명명하였다. 이 결과들은 가금류 및 사람에게 전파된 AIV H5N1아형의 진단에 적용될 수 있을 뿐 아니라, PCR이 사용되는 다른 신속검색법에도 널리 적용 될 수 있을 것으로 기대한다. The most rapid Real-time PCR based detection method for Avian influenza A virus (AIV) subtype H5N1 was developed. The target DNA sequence in this study was deduced from H5N1 subtype-specific 387 bp partial gene of hemagglutinin, and was synthesized by using PCR-based gene synthesis on the ground of safety. Real-Time PCR was performed by $GenSpector^{TM}$ using microchip-based, total $1{\mu}l$ of reaction mixture with extremely short time in each steps in PCR. The detection including PCR-amplication and analysis of melting temperature was totally completed within 13 min. The H5N1-specific 189 bp PCR product was correctly amplified until 2.4 molecules of hemagglutinin gene as minimum of templates. This kind of PCR was designated as Quick Real-Time PCR in this study and it could be applied to detect not only AIV H5N1, but also other pathogens using PCR-based detection.
버들치(Rhynchocypris oxycephalus)의 vitellogenin 생성량 측정을 이용한 수중 estrogen휴 화합물의 정량분석법에 관한 연구
김을환,이도부,한상훈,윤병수 경기대학교 부설 기초과학연구소 2005 경기대학교 기초과학논문집 Vol.18 No.1
Vitellogenin(Vtg), a phospholipoglycoprotein precursor of egg yolk, is synthesized and secreted from the liver in response to estrogens in female fish. Vtg is normally undetectable in the blood of male fish, but can be induced by exposure to chemicals possessing estrogenic activity. Thus, the presence of Vtg in blood of male fish can serve as a useful biomarker for assessing previous exposure to estrogenic compounds. In the present study, Vtg was abnormally expressed in Rhynchocypris oxycephalus treatment of Estradiol Benzoate (E_(2)). As the result, it was found that the level of Vtg in blood from R. oxycephalus was increased by treated quantity of E_(2) with dose-effect manner.