Escherichia coli에서 효모 Autonomously, Replicating Sequence Binding Factor 1의 발현과 정제

조기남,김지영,노현모,정구흥 경희대학교 유전공학연구소 1993 遺傳工學論文集 Vol.5 No.-

Autonomousely replicating sequence Binding Factor 1(ABF1) is a DNA-binding protein that specifically recognizes the RTCRYN5ACG at many sites in the yeast genome including the promoter element, mating-type silencer and ARSs. To express the intact full-length ABF1 gene in E. coli, the ABF1 gene has been cloned into pMAL-c2. Optimal expression of the fusion protein of ABF1 and MBP(maltose binding protein) was obtained by IPTG(Isoprophyl-β-D-thiogalactopyranoside) induction at 5 hr at 32℃. ABF1 fusion protein was purified by amylose resin affinity chromatography. Fusion protein of MBP and ABF1 was digested by Factor Xa and characterized by gel retardation assay.

효모 ABF1 단백질의 DNA Binding 부위에 대한 구조 기능 연구

조기남,이상경,김홍태,김지영,노현모,전구홍,Cho, Gi-Nam,Lee, Sang-Kyung,Kim, Hong-Tae,Kim, Ji-Young,Rho, Hyune-Mo,Jung, Gu-Hung 한국미생물학회 1994 미생물학회지 Vol.32 No.2

ABF1(Autonomously replicating sequence Binding Factor 1)은 효모 genome에서 $RTCRYN_5ACG$의 염기 서열을 가지고 있는 promoter, mating-type silencer, ARS에 결합하는 DNA binding 단백질이다. E. coli 에서 ABF1 유전자를 발현하기 위하여, ABF1 유전자를 pMAL-c2 벡터에 cloning하였다.(pMAHW). pMAHW를 E. coli에 형질전환하여, ABF1 융합단백질을 발현시키고, amylose resin affinity chromatography에 의하여 분리하였다. Factor Xa protease를 이용하여 분리된 융합단백질로부터 maltose binding protein을 잘라낸 후에 gel retardation analysis 방법으로 분리된 ABF1이 ARS1에 결합하는 능력을 지니고 있음을 확인하였다. DNA 결합에 관련된 부위를 찾기 위하여, 비전형적인 zinc finger motif가 위치하는 자리에서 pMAHW의 ABF1 유전자에 His-61을 다른 아미노산으로 치환하였다. DNA binding 부위로 추정되는 ABF1 단백질의 중간지역에 Leu-353, Leu-360를 다른 아미노산으로 치환하였다. Site-specific mutagenesis 를 통해 만들어진 mutant를 gel retardation analysis와 complementation test를 통해서 비전형적인 zinc finger motif이외에 다른 DNA binding motif가 있는 것을 알 수 있었다. Autonomously replicating sequence Binding Factor 1(ABF1) is a DNA-binding protein that specifically recognizes the $RTCRYN_5ACG$ at many sites in the yeast genome including the promoter element, mating-type silencer and ARS. To express the intact full-length ABF1 gene in E. coli, the ABF1 gene has been cloned into pMAL-c2 and His-61, Leu-353 and Leu-360 were substituted with other amino acid. ABF1 fusion proteins of wild type ABF1 and H61A, L353R and L360R nutants were purified by amylose resin affinity chromatography. Fusion protein of MBP and ABF1 was digested by Factor Xa and Characterized by gel retardation assay and complementation test. As aresult, we suggested that other DNA binding motif except atypical inc-finger motif is in the middle region of ABF1.

Contribution of HANARO Irradiation Technologies to National Nuclear R&D

조기남,조만순,양성우,박상준 한국원자력학회 2014 Nuclear Engineering and Technology Vol.46 No.4

HANARO is a multipurpose research reactor located at the Korea Atomic Energy Research Institute (KAERI). Since thecommencement of its operation in 1995, various neutron irradiation facilities, such as rabbit irradiation facilities, fuel test loop(FTL) facilities, capsule irradiation facilities, and neutron transmutation doping (NTD) facilities, have been developed andactively utilized for various nuclear material irradiation tests requested by users from research institutes, universities, andindustries. Most irradiation tests have been related to national R&D relevant to present nuclear power reactors such as theageing management and safety evaluation of the components. Based on the accumulated experience as well as thesophisticated requirements of users, HANARO has recently supported national R&D projects relevant to new nuclear systemsincluding the System-integrated Modular Advanced Reactor (SMART), research reactors, and future nuclear systems. Thispaper documents the current state and utilization of irradiation facilities in HANARO, and summarizes ongoing researchefforts to deploy advanced irradiation technology.

심뇌통(心腦通),강지연수단(降脂延壽丹) 및 청심조기탕(淸心調氣湯)의 고지혈증(高脂血症)에 대(對)한 실험적(實驗的) 연구(硏究)

김영석,문상관,고창남,조기호,배형섭,이경섭,노상겸,조효란,곽위성,Kim, Young-Suk,Moon, Sang-Kwan,Ko, Chang-Nam,Joh, Ki-Ho,Bae, Hyung-Sup,Lee, Kyung-Sup,Bao, Pei-Rong,Cao, Xiao-Lan 대한한방내과학회 1998 大韓韓方內科學會誌 Vol.19 No.1

In order to evaluate the effects of SIMNETONG, GANGJIYOUNSOODAN and CHUNGSIMJOKITANG(Xinnao-Tong, Jiangzhiyanshou-Dan and Qingxintiaoqi-Tang) on hyperlipidemia, the experimental study was performed in rats without or with hyperlipidemia induced by 2% cholesterol fed-diet or Triton WR-1339. And the levels of serum total cholesterol, triglyceride, HDL-cholesterol, LDL-cholesterol and phospholipid were measured. The results are summarized as follows : In the model of hyperlipidemia induced by 2% cholesterol Fed-diet 1. Both low & highly concentrated GANGJIYOUNSOODAN showed significantly decreasing effects on the level of serum total cholesterol while SIMNETONG & CHUNGSIMJOKlTANG didn't. 2. GANGJIYOUNSOODAN, SIMINETONG and CHUNGSIMJOKITANG showed decreasing effects on the level of serum triglyceride with no statistical significance. 3. Only highly concentrated GANGJIYOUNSOODAN showed increasing effects on the level of serum HDL-cholesterol while the others had no effects. 4. Only highly concentrated GANGJIYOUNSOODAN had decreasing effects on the level of serum LDL-cholesterol while the others had no effects. 5. Highly concentrated GANGJIYOUNSOODAN and CHUNGSIMJOKITANG showed significantly decreasing effects on the level of serum phospholipid while the other had no effects. In the model of hyperlipidemia induced by Triton WR-1339 6. SIMNETONG, GANGJIYOUNSOODAN and CHUNGSIMJOKITANG showed slightly decreasing effects on the levels of serum total cholesterol and triglyceride with no statistical significance. 7. None of SIMNETONG, GANGJIYOUNSOODAN and CHUNGSIMJOKITANG showed any effects on the level of serum HDL-cholesterol. 8. Highly concentrated GANGJIYOUNSOODAN and CHUNGSIMJOKITANG showed decreasing effects on the level of serum LDL-cholesterol but the other had no effects. 9. All of highly concentrated SIMNETONG, GANGJIYOUNSOODAN and CHUNGSIMJOKITANG showed decreasing effects on the level of serum phospholipid with statistical significance. These results show that GANGJIYOUNSOODAN and CHUNGSIMJOKITANG, in orders, have antihyperlipidemic effects. Further study on the mechanism and on the herb screening of these prescription should be followed.

급성기 말초성 안면신경마비에 대한 한방치료와 한양방 병용치료의 효과비교

조기호(Ki-Ho Cho),정우상(Woo-Sang Jung),홍진우(Jin-Woo Hong),황재웅(Jae-Woong Hwang),나병조(Byung-Jo Na),박성욱(Seong-Uk Park),문상관(Sang-Kwan Moon),박정미(Jung-Mi Park),고창남(Chang-Nam Ko),김영석(Young-Suk Kim),배형섭(Hyung-Sup Ba 대한한의학회 2008 대한한의학회지 Vol.29 No.1

뇌졸중 환자의 견관절 통증에 대한 봉독약침의 임상효과

고창남(Chang-nam Ko),민인규(In-kyu Min),박성욱(Sung-wook Park),정우상(Woo-sang Jung),문상관(Sang-kwan Moon),박정미(Jung-mi Park),조기호(Ki-ho Cho),김영석(Young-suk Kim),배형섭(Hyung-sup Bae) 대한한의학회 2007 대한한의학회지 Vol.28 No.1

A Preliminary Study on the Inhibitory Effect of Chunghyul-dan on Stroke Recurrence in Patients with Small Vessel Disease

조기호,지남규,정우상,박성욱,Sang-kwan Moon,고창남,Young-suk Kim,Hyung-sup Bae 대한한의학회 2007 대한한의학회지 Vol.28 No.1

Background: Chunghyul-dan is a combinatorial herbal medicine; previous studies reported it had therapeutic effects for microangiopathy, a major part in the progression of small vessel disease, as well as having anti-hypertensive, anti-hyperlipidemic, anti-apoptotic, anti-oxidative, and anti-inflammatory activities. Therefore, we examined the inhibitory effect of Chunghyul-dan on stroke recurrence in patients with small vessel disease. Methods: We prescribed Chunghyul-dan at 600 mg a day to patients with small vessel disease, and monitored stroke recurrence, drug compliances, and adverse effect for 1 year. We then performed follow-up brain MRI to find new vascular lesions after 1 year of Chunghyul-dan medication. For the subjects lost to follow-up, we assessed their prognosis after 1 year by telephone. Results: There were 73 subjects treated with Chunghyul-dan for 1 year; new vascular events were found in 3. Of the 85 subjects lost to follow-up, fifty four could be contacted, and eight of them had stroke recurrence. One year of Chunghyul-dan medication reduced the odds ratio of stroke recurrence by 75% compared to the subjects lost to follow-up and the rate increased to 88%, when adjusted for other relevant risk factors for stroke occurrence. These reductions were much higher than those of aspirin and other kinds of conventional anti-platelets. There was no adverse effect in any of the study subjects. Conclusions: We suggest Chunghyul-dan could be useful for inhibition of stroke recurrence. Further study with a randomized controlled trial is needed to confirm this suggestion.

Clinical Assessment of Usefulness,Effectiveness and Safety of Kyejibokryung-hwan on Various Neuropathies

고창남,박승욱,정우상,김윤경,문상관,김영숙,배형섭,조기호 대한한의학회 2006 대한한의학회지 Vol.27 No.4

We were to assess clinical effectiveness, safety and usefulness of Kyejibokryung-hwan (KBH), which has been used for Er-hyul similar with vascular or neurologic disorders causing sensory or motor abnormalities. This study is a retrospective single case series. Two hundred ninety five patients were treated with KBH for various neuropathies in our hospital. Of them, 120 cases were excluded because of insufficient medical records or failure to follow up; the remaining 175 were included in the analysis. There were 18 patients with sensory abnormalities and 28 with motor abnormalities after stroke. Of peripheral type, there were 14 with diabetic neuropathy, 12 with carpal tunnel syndrome, 41 with spinal radiculopathies, and 62 with idiopathic neuropathies, for which the effectiveness was assessed as 55.6%, 21.4%, 64.3%, 83.3%, 45.0%, and 56.5%, respectively. Adverse effects including indigestion or diarrhea were seen in 3.4% of the total patients. Taking the effectiveness and the safety together into consideration, the usefulness was assessed as 55.6%, 21.4%, 64.3%, 83.3%, 45.0%, and 54.8% for treating post-stroke sensory and motor abnormalities, diabetic neuropathy, carpal tunnel syndrome, spinal radiculopathies, and idiopathic neuropathies, respectively. In conclusion, we suggest that KBH is a useful herbal medicine for various neuropathies, especially of sensory type. We were to assess clinical effectiveness, safety and usefulness of Kyejibokryung-hwan (KBH), which has been used for Er-hyul similar with vascular or neurologic disorders causing sensory or motor abnormalities. This study is a retrospective single case series. Two hundred ninety five patients were treated with KBH for various neuropathies in our hospital. Of them, 120 cases were excluded because of insufficient medical records or failure to follow up; the remaining 175 were included in the analysis. There were 18 patients with sensory abnormalities and 28 with motor abnormalities after stroke. Of peripheral type, there were 14 with diabetic neuropathy, 12 with carpal tunnel syndrome, 41 with spinal radiculopathies, and 62 with idiopathic neuropathies, for which the effectiveness was assessed as 55.6%, 21.4%, 64.3%, 83.3%, 45.0%, and 56.5%, respectively. Adverse effects including indigestion or diarrhea were seen in 3.4% of the total patients. Taking the effectiveness and the safety together into consideration, the usefulness was assessed as 55.6%, 21.4%, 64.3%, 83.3%, 45.0%, and 54.8% for treating post-stroke sensory and motor abnormalities, diabetic neuropathy, carpal tunnel syndrome, spinal radiculopathies, and idiopathic neuropathies, respectively. In conclusion, we suggest that KBH is a useful herbal medicine for various neuropathies, especially of sensory type.

Effects of the Chungshimbohyeol-tang on the Gene Expression in Atherosclerotic Lesions

Ko,Chang-Nam,Kim,Young-Suk,Song,Mi-Deok,Cho,Ki-Ho,Lee,Kyung-Sup,Moon,Sang-Kwan,Bae,Hyung-Sup 慶熙大學校韓醫科大學韓醫學硏究所 1999 慶熙韓醫大論文集 Vol.22 No.1

The effects of the Chungshimbohyeol-tang on the expression of the MCP-1 and tissue factor genes which are important in the development of atherosclerotic lesions were investigated in the THP-1 human monocyte cell line using reverse transcription- polymerase chain reaction (RT-PCR). Treatment of the THP-1 cells with LPS and the Chungshimbohyeol-tang for 18 hr decreased the expressions of both MCP-1 and tissue factor mRNA in a dose dependent manner. Since it has been reported that NF-kB and AP-1, nuclear transcription factors, are involved in the transcriptional regulation of the MCP-1 and tissue factor gene, effects of the Chungshimbohyeol-tang on the activities of NF-kB and AP-1 were examined using a transient transfection assay and electrophoretic mobility shift assay(EMSA). When the THP-1 cells were transiently transfected with the plasmid containing 3 copies of the NF-kB binding site and the chloramphenicol acetyltransferase (CAT) gene, CAT activity was inhibited by the Chungshimbohyeol-tang. Treatment of the Chungshimbohyeol-tang inhibited protein/DNA binding of NF-kB to its cognate site as measured by EMSA. The Chungshimbohyeol-tang also inhibited AP-1 activation and its binding to DNA. Nuclear concentrations of NF-kB components and AP-1 proteins were also decreased by the treatment of the Chungshimbohyeol-tang. These results suggest that the Chungshimbohyeol-tang could prevent atherosclerosis through the down regulation of the MCP-1 and tissue factor gene expression which might be mediated by the inhibition of NF-kB and AP-1 activities and their binding to DNA.

Effects of Chungshimgunbi-tang on the Expressions of Tissue Factor and the Plasminogen Activator Inhibitor-1 Gene

Ko,Chang-Nam,Cho,Ki-Ho,Kim,Hong-Soon,Kim,Young-Suk,Lee,Kyung-Sup,Moon,Sang-Kwan,Bae,Hyung-Sup 慶熙大學校韓醫科大學韓醫學硏究所 1999 慶熙韓醫大論文集 Vol.22 No.1

Tissue factor와 PAI-1 유전자는 혈전증에 있어서 중요한 작용을 한다고 알려져 있다. 혈전증의 원인에 대해서 아직 자세히 알려져 있지는 않지만 protease 들의 응고 cascade의 일차적인 inhibitor이며 세포표면의 plama factor Ⅶ/Ⅶa의 receptor 및 co-factor로 알려진 tissue factor와 plasminogen activator를 저해하여 fibrin clot의 제거를 방해하는 것으로 알려진 plasminogen activator inhibitor-1 (PAI-1)이 관여하고 있다고 보고되고 있다. 이에 따라 본 연구에서는 한의학 임상에서 항혈전증에 빈용되는 청심건비탕이 Tissue factor와 PAI-1 유전자에 미치는 영향을 RT-PCR 방법을 사용하여 연구하였으며 이와 동시에 항혈전증이 기대되고 있는 가미형지탕, 가미이기탕, 가미정기산 및 가미소척탕등의 4개 처방도 screen하였다. 또한 그 세부방법으로 Tissue factor 유전자의 발현에 중요한 전사조절제인 NF-??B와 PAI-1 유전자의 발현에 중요한 AP-1 의 활성도를 chloramphenicol acetyltransferase (CAT) assay로 검증하였으며, Electrophoretic mobility shift assay로 NF-kB와 AP-1의 DNA와 결합을 조사하였다. 이상의 방법으로 청심건비탕이 혈전증 유발에 중요하다고 알려진 tissue factor와 plasminogen activator inhibitor-1 (PAI-1) 유전자의 발현에 대한 영향을 검토한 바 다음과 같은 결론을 얻었다. 1. 청심건비탕은 인간 monocyte cell line U937에서 tissue factor 와 PAI-1 mRNA의 생성을 농도 의존적으로 감소시켰으며 다른 약재들은 큰 차이를 보여주지 않았다. 2. NF-kB와 AP-1 의 활성도를 chloramphenicol acetyltransferase (CAT) assay로 검증한 결과 청심건비탕은 이들의 활성도를 유의하게 감소시켰다. 3. Electrophoretic mobility shift assay로 NF-kB와 AP-1의 DNA와 결합을 검증한 결과 청심건비탕은 이들 전사촉진제들이 DNA와 결합하는 것을 저해시킴이 규명되었다.