Solid phase Immunoassay using a Flow Cytometer : Quantitative and Qualitative Determination of Protein Antigens and a Hapten

김길현,한미영,윤도영,조보연,최명자,최인성,정태화 생화학분자생물학회 1991 생화학분자생물학회 춘계학술발표논문집 Vol.- No.-

Studies on Soybean Lipoxygenase 2

김길현,변시명,Kim, Kil-Hyun,Byun, Si-Myung 생화학분자생물학회 1981 한국생화학회지 Vol.14 No.2

Lipoxygenase 2 (E.C. 1. 13. 11. 12)를 대두로부터 친화성 크로마토그라피 방법과 DEAE-cellulose chromatography 방법으로 분리, 정제하였다. 친화성 크로마토그라파에서는 흡착제로서 linoleate-aminohexyl Sepharose 4B 를 사용하였으며, pH 6.8 에서 시행한 결과 lipoxygenase 2는 6배 정제되었고 수율은 91%였으나 lipoxygenase 1은 정제도가 오히려 떨어졌으며 수율은 15%이었다. 최종 lipoxygenase 2는 32.4 배 정제되었으며 수율은 28.5%이었다. 여기서 얻은 효소는 전기 영동법에 의해 거의 단일 단백질로 되어있음을 알 수 있었다. 여기서 얻은 lipoxygenase 2를 arachidonic acid와 반응 시킨후 sodium dithionite 로 환원시킨 다음 에테르로 추출한 물질중에 prostaglandin $F_{2{\alpha}}$로 보이는 소량의 물질이 포함되어 있음을 TLC 방법에 의해 확인하였다. Soybean lipoxygenase 2 was purified by the combination of ammonium sulfate fractionation, affinity chromatography on linoleate-aminohexyl Sepharose 4B and DEAE-cellulose chromatography. Purification of lipoxygenase 2 through affinity chromatography resulted in 6-fold purification and the yield of 91%, while lipoxygenase 1 was retrogressed and recovery was only 15%. Overall purification of lipoxygenase 2 was 32.4 folds and the yield was 28.5%, respectively. The purified enzyme preparation was almost homogeneous electrophoretically. When the lipoxygenase 2 preparation was incubated with arachidonic acid, and subsequently reduced with sodium dithionite, a material from ether extract of the reaction mixture seemed to have some of prostaglandin $F_{2{\alpha}}$, which was identified through thin layer chromatography.

회원투고 : 보다 빠른 지적민원 (地籍民願)의 처리방안

김길현 대한지방행정공제회 1972 地方行政 Vol.21 No.226

회원투고작품 : 시군단위의 농업직급 페지

김길현 대한지방행정공제회 1971 地方行政 Vol.20 No.208

T lymphocyte에 관한 연구동향

김길현 한국미생물학회 1991 微生物과 産業 Vol.17 No.1

이 글에서는 다음의 내용을 다루었다. 1. T세포에 있어서 피아식별의 기본 메카니즘 2. T세포의 활성화와 그에 관련된 세포 표면 분자들 3. T세포가 분비하는 lymphokine

결핵성 담관염과 림프절염에 의한 진행성 담관협착을 수술로 치료한 1예

김길현,구양서,김건국,김현옥,김금하,고광일,정낙소,유상균,박동균,권광안,김연석,김유경,김주현 대한소화기내시경학회 2007 Clinical Endoscopy Vol.35 No.4

Obstructive jaundice is most commonly attributed to a malignancy or stones affecting the common bile duct. Biliary tuberculosis and lymphadenitis around the periportal area have also been implicated but cases are quite rare. A 24 year old man presented with jaundice and abdominal pain for 3 days. Abdominal CT and ERCP revealed a stricture of the extrahepatic bile duct with multiple enlarged lymph nodes showing necrotic foci located at the periportal area. The colonoscopic biopsy showed evidence of M. tuberculosis. The patient was treated with ERBD insertion and oral anti-tuberculosis therapy. However, the abdominal pain recurred and there was progressive stenosis of the common bile duct. A bile duct resection with choledochojejunostomy was subsequently performed. Frozen sections revealed granulomatous inflammation with caseation necrosis, which was consistent with tuberculosis. We report a case of tuberculous cholangitis and lymphadenitis with obstructive jaundice that was managed surgically due to the progressive stricture of the bile duct. (Korean J Gastrointest Endosc 2007;35:287-291) 폐쇄성 황달을 일으키는 흔한 원인은 담도결석과 췌담도 종양이나, 간담도 결핵 또는 간문맥의 결핵성 림프절염에 의한 경우는 매우 드물다. 최근 3일 전부터 발생한 황달을 주소로 내원한 24세 남자 환자에서, 복부 전산화 단층촬영과 내시경 역행성 담췌관조영술(ERCP) 검사에서 총담관의 협착과 간문맥 부위에 다수의 림프절 종대 소견을 보이고, 대장 내시경검사에서 동반된 장결핵이 있어, 결핵성 림프절염으로 진단하고 내시경 역행성 담즙배액술 및 항결핵제 복용 등 내과적 치료를 시행하였다. 그러나 내과적 치료에도 불구하고 담관협착이 진행되어 담관 절제 및 총담관공장문합술을 시행하였다. 수술검체의 조직검사는 담관염 및 주위 림프절에 건락괴사를 동반한 만성육아종성 염증소견을 보여 결핵성 담관염 및 림프절염으로 확진하였다. 이에 결핵성 담관염 및 림프절염에 의해 담관협착이 진행되어 수술로 치료한 1예를 보고하는 바이다.

Telomerase Activity is Constitutively Expressed in the Murine CD8+ T Cells and Controlled Transcriptionally and Post-Translationally

김길현,김소정,김미형 대한면역학회 2004 Immune Network Vol.4 No.3

Background: Telomerase, a ribonucleoprotein enzyme capable of synthesizing telomeric repeats, attracts attention for its possible role in determining the replicative capacity of normal somatic cells, transformed cells, and cells of the germline lineage. Differently from normal somatic cells with no telomerase activity, normal lymphocytes has been reported to have telomerase activity comparable to that found in transformed cells during development and activation, which substantiate a role in supporting the capacity of lymphocytes for extensive clonal expansion. Methods: Here, in order to define the telomerase regulation in murine T lymphocytes, telomerase activity in cloned murine CD8 T cells and na ve CD8 T cells isolated from C57BL/6 mice was examined. Next, the regulatory mechanism of telomerase activity at transcriptional and post- translational levels was investigated by determining the expression level of the TERT protein, a key component for telomerase activity. Results: It was demonstrated that telomerase activity was expressed in an inactivated state as well as in an activated state in the murine CD8 T lymphocytes by using TRAP assay. The increase of telomerase activity was partially dependent on the net increase of TERT expression. Also, telomerase activity was decreased after treatment with protein kinase inhibitors, indicating that telomerase activation was prevented by inhibition of phosphorylation. Conclusion: Therefore, these results suggest that telomerase activity is constitutively expressed in the murine resting T lymphocytes and controlled by both transcriptional regulation and post-translational modifications. (Immune Network 2004;4(3):166-175)

대두 Lipoxygenase 2 에 관한 연구

김길현,변시명 ( Kil - Hyun Kim,Si Myung Byun ) 생화학분자생물학회 1981 BMB Reports Vol.14 No.2

Soybean lipoxygenase 2 was purified by the combination of ammonium sulfate fractionation, affinity chromatography on linoleate-aminohexyl Sepharose 4 B and DEAE-cellulose chromatography. Purification of lipoxygenase 2 through affinity chromatography resulted in 6-fold purification and the yield of 91%, while lipoxygenase 1 was retrogressed and recovery was only 15%. Overall purification of lipoxygenase 2 was 32.4 folds and the yield was 28.5%, respectively. The purified enzyme preparation was almost homogeneous electrophoretically. When the lipoxygenase 2 preparation was incubated with arachidonic acid, and subsequently reduced with sodium dithionite, a material from ether extract of the reaction mixture seemed to have some of prostaglandin F_(2α), which was identified through thin layer chromatography.

Optimization of a Virus-Induced Gene Silencing System with Soybean yellow common mosaic virus for Gene Function Studies in Soybeans

김길현,임승모,강양재,윤민영,남문,전태환,서민정,백성범,이점호,문중경,이석하,이수헌,임현섭,문재선,박창환 한국식물병리학회 2016 Plant Pathology Journal Vol.32 No.2

Virus-induced gene silencing (VIGS) is an effective toolfor the study of soybean gene function. Successful VIGSdepends on the interaction between virus spread andplant growth, which can be influenced by environmentalconditions. Recently, we developed a new VIGS systemderived from the Soybean yellow common mosaicvirus (SYCMV). Here, we investigated several environmentaland developmental factors to improve the efficiencyof a SYCMV-based VIGS system to optimize thefunctional analysis of the soybean. Following SYCMV:Glycine max-phytoene desaturase (GmPDS) infiltration,we investigated the effect of photoperiod, inoculationtime, concentration of Agrobacterium inoculm, andgrowth temperature on VIGS efficiency. In addition,the relative expression of GmPDS between non-silencedand silenced plants was measured by qRT-PCR. Wefound that gene silencing efficiency was highest at aphotoperiod of 16/8 h (light/dark) at a growth temperatureof approximately 27°C following syringe infiltrationto unrolled unifoliolate leaves in cotyledon stagewith a final SYCMV:GmPDS optimal density (OD)600 of2.0. Using this optimized protocol, we achieved high efficiencyof GmPDS-silencing in various soybean germplasmsincluding cultivated and wild soybeans. Wealso confirmed that VIGS occurred in the entire plant,including the root, stem, leaves, and flowers, and couldtransmit GmPDS to other soybean germplasms viamechanical inoculation. This optimized protocol usinga SYCMV-based VIGS system in the soybean shouldprovide a fast and effective method to elucidate genefunctions and for use in large-scale screening experiments

스포츠센터 이용에 관한 소비자 태도분석

김길현,이규희 대구산업정보대학 2000 논문집 Vol.14 No.1

The sports center has received much attention in recent years as a result of rapid economic growth and elevation of standards of living. The purpose of this study is to propose a scheme which is necessary to arouse consumer satisfaction through the attitude test concerning sports center. A field survey was conducted to study the attitude of consumer in Gangsan sports center, one of the largest sports center in Daegu area. The main results are as follows: 1. facilities which consumers used most frequently are swimming pool and training field that are able to use alone. 2. the major factors influencing sports center are the quality and variety of facilities. 3. the main purposes of using sports center are for maintaining or promoting health and for relieving stress. 4. the result factors of using sports center show significant difference according to demographic variables.