미공개정보이용행위 입증에 관한 연구 - 실증적 분석을 통한 미공개정보이용행위 규제의 실효성 강화방안 -

이곤호(Lee, Gon Ho) 한국증권법학회 2019 증권법연구 Vol.20 No.2

Reprogramming of the Activity of the Activator/Dissociation Transposon Family during Plant Regeneration in Rice

김철민,제병일,Hai Long Piao,Soon Ju Park,김민정,Sung Han Park,Jin Young Park,Su Hyun Park,Eun Kyeong Lee,Nam Soo Chon,Yong Jae Won,Gi Hwan Lee,Min Hee Nam,Doh Won Yun,Myung Chul Lee,Young Soon Cha,이곤호,은무영,한창덕 한국분자세포생물학회 2002 Molecules and cells Vol.14 No.2

Many aspects of epigenetic phenomena have been elu-cidated via studies of transposable elements. An active transposable element frequently loses its ability to mobilize and goes into an inactive state during devel-opment. In this study, we describe the cyclic activity of a maize transposable element dissociation (Ds) in rice. In rice genome, Ds undergoes the spontaneous loss of mobility. However, an inactive state of Ds can be changed into an active state during tissue culture. The recovery of mobility accompanies not only changes in the methylation patterns of the terminal region of Ds, but also alteration in the steady state level of the acti-vator (Ac) mRNA that is expressed by a constitutive CaMV 35S promoter. Furthermore, the Ds-reactiva-tion process is not random, but stage-specific during plantlet regeneration. Our findings have expanded previous observations on Ac reactivation in the tissue culture of maize.

Pichia pastoris로부터 Toll-like Receptor 9의 세포 내 도메인 단백질의 발현과 순수분리 정제

이균영,이곤호,Lee Kyun-Young,Lee Kon-Ho 한국식물생명공학회 2005 식물생명공학회지 Vol.32 No.4

Methylotrophic 효모 Pichia pastoris 발현시스템을 사용하여 인간 TLR9 단백질의 세포내 TIR 도메인을 발현하였다. TIR 단백질이 P. pastoris에서 발현되어 배지 속으로 분비되는 것을 SDS-PAGE로 확인하였고, 발현된 단백질을 western-blot, MALDI-TOF 질량분석으로 동정하였다. 이를 통하여 TIR 딘백질이 P. pastoris에서 안정적으로 발현됨을 알 수 있었다. 그리고 발현된 단백질을 니켈 친화, 양이온교환수지, 겔 투과 크로마토그라피를 사용하여 순수 분리 정제하였다. P. pastoris를 이용한 단백질의 발현과 정제방법은 대장균에서 잘 발현되지 않는 단백질의 발현에 응용될 수 있을 것이다. Toll-like receptors (TLR) are important components of innate immunity in the defense against pathogens. TLRs recognize pathogen-associated common molecular patterns. TLRs are similar to the receptors involved in defense responses in plants. TLR protein is a type 1 membrane protein, consisting of an extracellular domain containing leucine-rich repeats and a cytoplasmic domain. The cytoplasmic domain delivers ligand recognition signals that result in production of anti-microbial agents. The cytoplasmic domain (amino acid 858-1032) of toll-like receptor 9 has been expressed using methylotrophic yeast Pichia pastoris. The protein expression was confirmed by Western-blot, N-terminal sequencing and MALDl-TOF mass spectrometry. The proteins have been purified by nickel affinity, cation exchange and gel-filtration chromatography.

퉁퉁마디로부터 색소체 외막 단백질 유전자의 분리 및 발현분석

네티 엘마와티,차준영,양영실,정민희,신동진,이병현,이곤호,손대영,Ermawati Netty,Cha, Joon-Yung,Liang, Yingshi,Jung, Min-Hee,Shin, Dongjin,Lee, Byung-Hyun,Lee, Kon-Ho,Son, Daeyoung 한국식물생명공학회 2004 식물생명공학회지 Vol.31 No.4

Differential display 방법으로 NaCl에 의하여 발현이 증가되는 cDNA들을 분리하였으며 그중 하나가 식물유래의 outer envelope membrane protein과 높은 유사성을 보였으므로 이를 ShOEP로 명명하였다. ShOEP는 1293 bp 길이에 359개의 아미노산으로 구성된 open reading frame을 포함하고 있으며, 이로부터 추정되는 분자량은 8.9 kDa이었다. ShOEP 단백질은 애기장대의 OEP와는 40.6%, 시금치와는 38%의 유사성을 나타내었다. Northern 분석결과, ShOEP 유전자는 NaCl의 농도가 증가함에 따라 발현량이 급격히 증가하는 것으로 나타났다. 염생식물인 퉁퉁마디의 OEP는 PEG에 의하여 발현이 증가하는 반면 비염생식물인 애기장대의 OEP는 큰 차이를 보이지 않았다. 효모 complementation 실험결과 ShOEP는 NaCl에 특이적이었으며 식물의 염분스트레스 내성 기작에 직접적으로 관여하고 있음을 알 수 있었다. Complementary DNA encoding chloroplast outer envelope membrane protein (OEP) from the halophyte Salicornia herbacea has been cloned and sequenced. The full length cDNA is 596 bp and encodes a polypeptide of 91 amino acid residues with a molecular mass of 8.9 kDa. The expression level of ShOEP increased by salt, drought and ABA treatments. ShOEP expression was largely induced in roots and shoots by high salts. The biological function of ShOEP was examined by yeast complementation. ShOEP can suppress Na$^{+}$ sensitivity of yeast mutant (cnb$\Delta$) in the presence of salt. These results suggest that ShOEP is a salt inducible gene and may have functions in the regulation of plant salt stress.ant salt stress.

퉁퉁마디로부터 염에 의하여 유도되는 Aldolase 유전자의 분리 및 발현분석

차준영,네티엘마와티,김순길,이증주,임채오,정우식,이곤호,손대영,Cha, Joon-Yung,Netty Ermawati,Kim, Soon-Gil,Lee, Jeung-Joo,Lim, Chae-Oh,Chung, Woo-Sik,Lee, Kon-Ho,Son, Dae-Young 한국식물생명공학회 2003 식물생명공학회지 Vol.30 No.4

토양 내의 고농도의 염은 심각한 환경스트레스 중의 하나로 농작물의 생산을 감소시킨다. 식물은 염 스트레스로부터 벗어나기 위하여 많은 단백질을 합성한다든지 유전자들의 발현을 조절하는 등 여러 가지 생리, 생화학적인 변화를 일으킨다. 퉁퉁마디는 우리나라에 자생하는 염생식물로 갯벌과 염전주위에서 생육한다. 퉁퉁마디의 생화학적, 분자생물학적 내염성 기구를 이해하기 위하여 differential display방법으로 NaCl에 의하여 발현이 증가되는 cDNA들을 분리하였다. 본 연구에서는 그 중 하나인 ShADL의 특성을 조사하였다. ShADL은fructose-1, 6-bisphosphate aldolase와 높은 유사성을 보였다. 이 유전자는 1293bp길이에 359개의 아미노산으로 구성된 open reading frame을 포함하고 있으며, 이로부터 추정되는 분자량은 39 kDa이었다. ShADL단백질은 애기장대의 aldolase와 86%의 높은 유사성을 나타내었으며 같은 염생식물인 com-mon ice plant의 adolase와는 78%의 유사성을 보였다. Northern 분석결과, ShADL 유전자는 NaCl의 농도가 증가함에 따라 발현량이 급격히 증가하는 것으로 나타났다. Soil salinity is one of the most serious abiotic stresses limiting the productivity of agricultural crops. To cope with salt stress, plants respond with physiological, developmental and biochemical changes, including the synthesis of a number of proteins and the induction of gene expression. Salicornia herbacea is a halophytic plant that grows in salt marshes and on muddy seashores. In order to understand the biochemical and molecular mechanisms of salt tolerance in S. herbacea, we isolated several genes that involved in the salt tolerance by mRNA differential display. Here we report the cloning of a cDNA encoding fructose-1, 6-bisphosphate aldolase, named ShADL, which is 1293 bp long and contains an open reading frame consisted of 359 amino acids with calculated molecular mass of 39 kDa. ShADL protein showed 86% identity with Arabidopsis and 78% with aldolase of common ice plant. Northern blot analysis revealed that the transcript of ShADL gene was increased dramatically depending on the NaCl concentrations.

Characterization of Arabidopsis Histidine Kinase 3 and Proteomic Analysis of Its Mutant

양영실,차준영,네티 엘마와티,정민희,이곤호,손대영,Liang Ying-Shi,Cha Joon-Yung,Ermawati Netty,Jung Min-Hee,Lee Kon-Ho,Son Dae-Young Korean Society of Life Science 2006 생명과학회지 Vol.16 No.3

Histidine kinase는 식물의 신호전달기작에서 매우 중요한 역할을 한다. 본 연구에서는 애기장대 histidine kinase 3 (AHK3)의 식물체내에서의 기능을 조사하였으며 이 유전자의 결손 돌연변이체인 ahk3에 trans-zeatin (t-zeatin)을 처리하여 유전자와 단백질의 발현양상을 분석하였다. ahk3는 야생형 식물체에 비하여 캘러스 형성, 유모의 성장, 잎의 노화과정에서 t-zeatin에 대한 감수성이 줄어들었다. 프로테옴 분석 결과 eukaryotic translation initiation factor 5A-2, auxin binding glutathione S-transferase, NDPK1 등은 야생형의 애기장대에서는 t-zeatin에 의하여 발현이 증가하는 반면 ahk3에서는 증가하지 않는 것으로 나타났다. 또한 cytokinin처리에 의하여 발현이 증가하는 것으로 보고된 A-type response regulator들 중에서 ARR4와 ARR16의 발현양이 ahk3에서는 현저하게 감소하는 것으로 나타났다. 이러한 결과들은 AHK3가 cytonin신호전달기작에서 매우 중요한 역할을 하며, 프로테옴 분석에 의하여 동정된 단백질들과 ARR4, ARR16은 AHK3에 의해 매개되는 cytokinin 신호전달과정에서 중요한 역할을 할 것으로 생각된다. Histidine kinase plays important roles in signal transduction in plant. We characterized the function of Arabidopsis histidine kinase 3 (AHK3) and analyzed the expression patterns of genes and proteins in its mutant ahk3 by trans-zeatin (t-zeatin). The ahk3 exhibited decreased sensitivity to t-zeatin during callus formation, seedling growth, and leaf senescence. From proteomic analysis of ahk3, eukaryotic translation initiation factor 5A-2, auxin binding glutathione S-transferase, and NDPK1 were identified not to be induced by t-zeatin, when compared to the wild-type. In addition, the expression levels of ARR4 and ARR16 among A-type response regulators (ARRs) markedly decreased in ahk3 by t-zeatin treatment. These results suggest that AHK3 plays an important role in cytokinin signaling and the proteins identified from proteomic analysis and specific ARRs, ARR4 and ARR16 may be directly or indirectly associated in AHK3-mediated cytokinin signaling.

Pichia pastoris로부터 Toll-like Receptor 9의 세포 내 도메인 단백질의 발현과 순수분리 정제

이균영,이곤호 Plant molecular biology and biotechnology research 2005 Plant molecular biology and biotechnology research Vol.2005 No.

Toll-like receptors (TLR) are important components of innate immunity in the defense against pathogens. TLRs recognize pathogen-associated common molecular patterns. TLRs are similar to the receptors involved in defense responses in plants. TLR protein is a type 1 membrane protein, consisting of an extracellular domain containing leucine-rich repeats and a cytoplasmic domain. The cytoplasmic domain delivers ligand recognition signals that result in production of anti-microbial agents. The cytoplasmic domain (amino acid 858-1032) of toll-like receptor 9 has been expressed using methylotrophic yeast Pichia pastoris. The protein expression was confirmed by Western-blot, N-terminal sequencing and MALDI-TOF mass spectrometry. The proteins have been purified by nickel affinity, cation exchange and gel-filtration chromatography.

퉁퉁마디로부터 색소체 외막 단백질 유전자의 분리 및 발현분석

네티 엘마와티,차준영,양영실,정민희,신동진,이병현,이곤호,손대영 Plant molecular biology and biotechnology research 2004 Plant molecular biology and biotechnology research Vol.2004 No.-

Complementary DNA encoding chloroplast outer envelope membrane protein (OEP) from the halophyte Salicornia herbacea has been cloned and sequenced. The full length cDNA is 596 bp and encodes a polypeptide of 91 amino acid residues with a molecular mass of 8.9 kDa. The expression level of ShOEP increased by salt, drought and ABA treatments. ShOEP expression was largely induced in roots and shoots by high salts. The biological function of ShOEP was examined by yeast complementation. ShOEP can suppress Na+ sensitivity of yeast mutant (cnbΔ) in the presence of salt. These results suggest that ShOEP is a salt inducible gene and may have functions in the regulation of plant salt stress.

퉁퉁마디로부터 염에 의하여 유도되는 Aldolase 유전자의 분리 및 발현분석

차준영,네티 엘마와티,김순길,이증주,임채오,정우식,이곤호,손대영 Plant molecular biology and biotechnology research 2003 Plant molecular biology and biotechnology research Vol.2003 No.-

Soil salinity is one of the most serious abiotic stresses limiting the productivity of agricultural crops. To cope with salt stress, plants respond with physiological, developmental and biochemical changes, including the synthesis of a number of proteins and the induction of gene expression. Salicornia herbacea is a halophytic plant that grows in salt marches and on muddy seashores. In order to understand the biochemical and molecular mechanisms of salt tolerance in S. herbacea, we isolated several genes that involved in the salt tolerance by mRNA differential display.Here we report the cloning of a cDNA encoding fructose-1, 6-bisphosphate aldolase, named ShADL, which is 1293 bp long and contains an open reading frame consisted of 359 amino acids with calculated molecular mass of 39 kDa. ShADL protein showed 86% identity with Arabidopsis and 78% with aldolase of common ice plant. Northern blot analysis revealed that the transcript of ShADL gene was increased dramatically depending on the NaCI concentrations.