다중 중합효소 연쇄반응을 이용한 소의 Johne병 진단 기법 확립

김종배,송혜원,김근희,김홍,신광순,김두 대한의생명과학회 2000 Journal of biomedical laboratory sciences Vol.6 No.1

반추수에서 발생하는 Johne병의 조기 진단 방법을 제시하고 이 질병의 원인체와 미생물학적 특징이 유사한 M. bovis, M. avium 등의 mycobacteria감염증을 감별 진단하는 방법을 개발하기 위하여 Mycobacterium 균속의 표준균주를 사용하여 중합효소 연쇄반응을 확립하였다. Johne병으로 의심되는 소의 혈액과 유즙을 채취하여 분리한 단핵구 및 거식세포로부터 genomic DNA를 추출하였다. 각 시료로부터 추출한 DNA를 template로 이용하여 Mycobacterium spp.에 특이적인 16S rDNA primer set를 이용한 PCR을 수행하여 시료내의 mycobacterial DNA 보유 여부를 확인하였다. 한편 mycobacteria 양성으로 확인된 시료는 M. avium complex 균종에 특이한 16S rDNA 염기서열을 기초로하여 제작한 primer set와 M. paratuberculosis 의 IS900 sequence에 특이한 primer set를 이용하여 duplex PCR을 수행하여 Johne병 원인체의 보균 여부를 조사하였으며, 이 결과를 oligonucleotide probe를 사용한 Southern blot hybridization을 통하여 다시 확인하였다. 이와 같은 duplex PCR 기법을 실제 축산 현장에서 수집한 유즙과 말초혈액으로부터 분리한 단핵구 및 거식세포 시료에 적용한 결과 본 연구에서 확립한 duplex PCR기법 유용성을 확인할 수 있었다. Diagnosis of Bovine Johne's Disease Using Multiplex Polymerase Chain Reactions In order to improve the early diagnosis of Johne's disease in ruminants, duplex polymerase chain reaction system for the detection of the etiologic agent of M. paratuberculosis and for the differentiation of other mycobacterial animal pathogens, such as M. bovis and M. avium, was applied. Genomic DNAs were purified from peripheral blood monocytes or milk macrophages and were used as templates in the duplex PCR. Detection of Mycobacterium spp. in the specimen was carried out by PCR using primer set specific to the mycobacterial 16S rDNA. And then, mycobacterial DNA-positive specimens were further differentiated with duplex PCR system which was composed of primer sets specific to 16S rDNA of M. avium complex and IS900 gene of M. paratuberculosis. The results were re-confirmed by Southern blot hybridization with oligonucleotide specific to the internal sequence of IS900 PCR amplicons. The applicability of this duplex PCR system was evaluated with DNAs extracted from clinical specimens of peripheral blood monocytes and milk macrophages. In summary, the duplex PCR amplification system described in this experiment is promising molecular technique for the early diagnosis of Johne's disease in ruminants.

Monascus anka로부터 유기배양에 의한 적색소의 대량생산

김희구,박근태,손홍주 한국식품영양학회 1998 韓國食品營養學會誌 Vol.11 No.6

Monascus anka KFCC 4478의 적색소 생산력을 증가시키기 위하여 발효조 배양조건을 검토한 후, 탄소원을 연속적 또는 간헐적으로 공급하는 유기배양을 하였다. 5l 용량의 발효조를 이용하여 배양시 교반속도 200rpm, 통기량 1vvm 및 종균 접종량 10%(v/v)일 때 색소 생성량이 최대였다. 또한 벽면에 부착된 균사체를 재접종함으로써 색소 생성력이 향상되었다. Intermittent feeding fed-batch culture에 의하여 배양 9일만에 30g/l의 건조균체와 350(A_495)의 색소가 생성되었으며, continuous feeding fed-batch culture에 의하여 22g/l의 건조균체와 190(A_495)의 색소가 얻어졌다. 따라서 본 균주에 의하여 적색소를 생산코자 할 경우는 회분배양보다 유기배양이 더 우수한 것으로 나타났다. The production of red pigment from glucose by fed-batch culture of Monascus anka was investigated. In batch culture using fermentor, 200 rpm of agitation speed, 1vvm of aeration volume, and 10%(v/v) of inoculum size were optimal, respectively. The red pigment production was increased by removal of wall-attached mycelium. In an intermittent feeding fed-batch culture, dry cell weight increased to 30g/l, and the red pigment content reached 350 of absorbance at 495nm. In a continuous feeding fed-batch culture, dry cell weight increased to 22g/l, and the red pigment content reached 190 of absorbance at 495nm.

Monasucs anka의 적색조 생산 특성

김희구,박근태,손홍주 한국식품영양학회 1998 韓國食品營養學會誌 Vol.11 No.6

Monascus anka KFCC 4478의 적색소 생산력을 증가시키기 위하여 배지조성 및 배양조건에 대하여 검토하였다. 공시균주의 색소 생성력은 탄소원으로 glucose 4.0%, 유기질소원으로 peptone 3.0%, 무기질소원으로 NaNO_3 0.15%, 비타민 성분으로 thiamine-HCI 및 nicotinic acid 각 10㎍/l였으며, 배양온도 30℃, 초기 pH 7.0이었다. 특히 유기질소원과 무기질소원을 복합 처리함으로서 색소 생성력이 향상되었다. 결정된 색소생성 최적조건하에서 본 공시균주는 배양 5일만에 최대 균체 증식과 색소 생성력을 나타내었다. Optimal media and cultural conditions for the production of red pigment were established using Monascus anka KFCC 4478. The optimal temperature and initial pH for the production of red pigment were 30℃ and 7.0, respectively. Glucose turned out to be most suitable carbon source for red pigment production. Optimal glucose concentration was 3.0%. Addition combined of nitrogen sources of peptone and NaNO_3 induced good red pigment production. Thiamine-HCL and nicotinic acid were increased the production of red pigment. Under optimal conditions, maximum red pigment production and cell growth were observed after 5 days of incubation.

다결정 실리콘 태양전지의 상부 전극용 금속 박막 적용

김상수,임동건,이준신,심경석,김홍우,이만근 성균관대학교 1997 학술회의지원논문목록집 Vol.1997 No.-

We investigated grain boundary effect for terrestrial applications of solar cells with low cost, large area, and high efficiency. Grain boundaries are known as potential barriers and recombination centers for the photo generated charge carriers, which make it difficult to achieve a high efficiency cell. To reduce these effects of grain boundarues, we investigated various influencing factors such as thermal treatments various grid patterns selective wet etchings for grain boundarues, buried contact metallizations along grain boundaries, and use of metallic thin films. From the various grid patterns we learned that the series resistance of solar cell reduced open circuit voltage and consequently decreased the cell efficiency. This paper describes the effect of various grid patterns and the employment of metallic thin films for a top electrode.

토양에서 분리한 Toluene 분해 세균의 동정 및 일부 특성

김종구,박근태,이건,이상준,손홍주 부산대학교 환경문제연구소 1998 環境硏究報 Vol.16 No.-

경상남도 양산천 일대의 토양시료로 부터 toluene을 유일한 탄소원 및 에너지원으로 생육하는 미생물들을 분리하였고, 그 중 toluene의 분해능이 가장 우수한 균주를 선별하여 공시균으로 선택하였다. 공시균의 형태학적, 배양적, 생화학적 및 화학분류학적 제반 특성들을 토대로 분류학적 위치를 검토한 결과 Rhodococcus 속으로 동정되어 편의상 Rhodococcus sp. EL-43P로 명명하였다. Rhodococcus sp. EL-43P는 phenol 및 다양한 phenol 유도체, benzene 등의 방향족화합물 기질에서 생육할 수 있었다. 특히 phenol 분해에 관련된 각종 환경조건을 최적화시키면 실제의 phenol 함유 폐수처리공정에서 충분히 적용가능성이 있음을 알 수 있었다. Rhodococcus sp. EL-43P는 catechol 1,2-dioxygenase의 활성을 가지고 있어 ortho pathway에 의하여 phenol을 분해함을 알 수 있었다. 또한 본 공시균은 toluene dioxygenase의 활성도 가지고 있었다. The microorganisms which degrade toluene as the sole and energy source were isolated from soil around Yangsan river. Among them, the strain EL-43P had the highest toluene degradability and identified as a Rhodococcus sp. according to morphological, cultural, biochemical and chemotaxonomical properties. Rhodococcus sp. EL-43P was able to grow at the various chlorinated phenols such as o-, p-chlorophenol, 2,3-,2,5-, 2,6- and 3,4-dichlorophenol, and benzene, respectively. If detailed phenol degradation conditions are established, the treatment of phenol wastewater using Rhodococcus sp. EL-43P would be possible. Rhodococcus sp. EL-43P had the catechol 1,2-dioxygenase which degrades Phenol derivatives by ortho pathway. This strain also had toluene dioxygenase.

품종보호용 분자마커의 고추 내 도입

홍지영,최근원,김영채 경희대학교식량자원개발연구소 2006 硏究論文集 Vol.25 No.-

본 연구는 쥐의 adenosine deaminase(ADA) 유전자를 이용하여 개발된 고추 품종보호용 분자마커로서의 실용 가능성을 검토하기 위하여 실시되었다. 513bp 크기의 개발된 품종보호용 DNA 마커를 운반하는 A. tumefaciens LBA4404를 이용한 형질전환 실험에서 공동배양한 후 선발배지로 옮겨 배양한 '135Q' 와 '276F' 고추자엽 절편체는 두 계통 모두에서 84%와 80%의 높은 재분화율을 보였다. '135Q'와 '276F' 고추 자엽 절편체로부터 재분화된 신초들에 대한 분자마커 특히 primer를 이용한 PCR검정과 PCR로 확인된 잠정적인 형질전환 신초에 대한 Southern hybridization 분석 결과 품종보호용으로 개발된 513bp DNA 마커의 삽입을 확인하였다. PCR 검정 결과 '135Q'와 '276F' 고추 자엽 절편체로부터 재분화된 신초의 62.6%, 62.9%가 각각 형질전환체로 확인되었다. 이러한 안정적인 DNA 마커의 전이는 고추에서의 품종보호를 위한 활용 가능성을 제시한다. This study was carried out to investigate the possibility for the practical application of a molecular marker which was developed from mouse adenosine deaminase (ADA) gene for cultivar protection in hot pepper (Capsicum annuum L.). In the transformation of hot peppers using Agrobacterium tumefaciens LBA4404 carrying a 513 bp DNA marker for cultivar protection, the regeneration percentage from cotyledon explants of '135Q' and '276F' hot pepper lines after co-cultivation were 84 and 80%, respectively. Results of PCR analysis on regenerated shoots using marker-specific primers and Southern hybridization analysis on putative transgenic shoots confirmed by PCR indicated that the 513 bp DNA marker for cultivar protection was successfully transferred into the regenerated shoots. Twenty three and twenty four out of 33 and 37 regenerated shoots were transgenic in '135Q' and '276F' hot peppers, respectively. This stable transfer of the DNA marker suggested the possibility of practical application for cultivar protection in hot peppers.

모재의 경도가 R.F PACVD에 의하여 증착된 TiN의 경도와 밀착력에 미치는 영향(Ⅱ)

金鴻遇,朴達根 忠州大學校 1997 한국교통대학교 논문집 Vol.32 No.1

The goal of this study is to investigate the influence of mechnical property on the behavior of hardness and adhesion of TiN thin film deposited by the radio frequency plasma-assisted chemical vapor deposition after changing the hardness of substrate by heat treatment. X-ray diffraction analysis, RBS and EPMA spectra were used to investigate phase, structure and chemical composition of the deposited layer and the vickers hardness test and scratch adhesion test were performed to analyze microhardness and adhesion of the deposited layer. According to the results of this study, the substrate hardness does not change the chemical composition, stoichiometry and structure of the deposited TiN. ISE index is 1.96-1.99 for the substrate and 1.57-1.79 for the deposited TiN and the substrate hardness is in inverse proportion to the ISE index. While the deposited TiN has many cracks around the indentation for low substrate hardness, the number of cracks around the indentation decreased and the deformation is limited within the indentation. In measuring adhesion by SAT, the critical load generally increased as the substrate hardness decreased. Considering this result, the mechanical property of substrate as well as the deposited layer should be considered together and should be selected to fit the industrial applications.

랫드에서 법제유황과 비법제유황의 독성비교

김근수, 정유리, 이남진, 홍성희, 유종훈, 정재황, 강종구 충북대학교 동물의학연구소 2006 Journal of Biomedical and Translational Research Vol.7 No.2

본문참고

척추 수술시 자가혈액 수혈기를 이용한 자가수혈의 효과

김명호,유문집,유석주,박진영,정홍근,박희곤,문기혁 단국대학교 2000 論文集 Vol.35 No.-

Study Design : Spine fusion with fixation procedure is associated with substantial operative blood loss, which frequently necessitates transfusion. The authors evaluated the safety and efficacy of intraoperative autotransfusion using Cell Saver during spinal surgery. Summary of Literature Review : It has been reported that the contraindications to the use of intraoperative autologous transfusion are malignancy, infection, and gross wound contamination. Objectives : The purpose of this study was to evaluate the safety and efficacy of intraoperative autotransfusion using Cell Saver during spine surgery. Material and Methods : We retrospectively reviewed 62 patients who had operation for lumbosacral spinal stenosis and hypotensive anesthesia. Among them, Group Ⅰwhich received autotrans fusion using Cell Savers included 26 patients with fusion of 2 segments, and Group Ⅱ as control group included 13 patients. Group Ⅲ which received autotransfusion using Cell Savers included 17 patients with function of 3 segments, and Group Ⅳ as control group included 6 patients. We analyzed the amount of intraoperative blood salvage using a Cell Saver, the amount of total homologous transfusion amount, intraoperative blood loss, the amount of postoperative drainage amount and perioperative changes of hemoglobin, hematocrit and platelet count. Results : There was no statistical significance between Cell-Saver group and control group in amount of total homologous transfusion and intraoperative(except in 2 segment of fusion using cell saver: 86.5% increased) & postoperative blood loss(p>0.05). The circulatory balance(amount of intraoperative blood salvage using a Cell Saver plus amount of total homologous transfusion minus amount of total blood loss) was statistical significance between Cell-Saver group and Control group. The Cell-Saver group had positive balance and Control group had negative balance. There were no statistical differance in the evolusion of hemoglobin, hematoocrit and platelet count between Cell-Saver group and Control group(p>0.05). There was no transfusion reaction and thromboembolic disease in all group. Conclusion : In retrospective study, Cell Savers was safe and immediate availability in orthopedic surgery such as spinal surgery. It was effective methods for storing a sufficient volume of blood for spine surgery without affecting the patient's postoperative hemodynamic ststes, too. It was possible to operation without intraoperative homologous blood transfusion in 2 or 3 segment fusion. We may need further study.

역간접촉산화법에 의한 연안 하구 수질개선에 관한 연구

김원근,이찬원,전홍표 경남대학교 환경문제연구소 2002 환경연구 Vol.25 No.-

A field study was conducted to investigate the removal efficiency and performance of contact oxidation treatment system installed and operated in two locations of stream flowing to the Masan Bay. The removal of organic matters from stream water flowing to the already eutrophicated bay is getting more important in order to restore the water quality. The removal efficiencies of BOD, SS and ABS loading rate were 52~63%, 50~70% and 60~85%, respectively, whereas 10~25% of total nitrogen and 25~40% of total phosphorus were only removed. The effluent water quality did not meet the goal class of water regulation standards. Inflow rates to contact oxidation treatment showed seasonal and hourly variations. The treatment capacity installed is now enough to handle the present inflows, thus it was suggested that this system should be operated to supply only the volume of oxygen to require the actual loading to the treatment system and the aeration system would be intermittently operated for the improvement of nitrogen and phosphorus removal. It was found that inflow rate should be controlled for detention time of contact oxidation to maintain at least more than 1.5 hr. When microbiological species of sludge and sludge accumulation in the lower steps of contact oxidation process was examined, proper draw-out of sludge from the system was also an important operating parameter.