연구 보문 : 옥수수 B-peru 유전자가 도입된 담배 형질전환체의 페닐프로파노이드 생합성 유전자의 발현분석

임선형 ( Sun Hyung Lim ),하선화 ( Sun Hwa Ha ),김재광 ( Jae Kwang Kim ),손성한 ( Seong Han Sohn ),김동헌 ( Dong Hern Kim ),김정선 ( Jung Sun Kim ),김영미 ( Young Mi Kim ),이종열 ( Jong Ryeol Lee ) 한국국제농업개발학회 2010 韓國國際農業開發學會誌 Vol.22 No.1

벼 HMG-CoA 환원효소의 특성연구

하선화,김제현,백융기,김종범,김종국,황영수 ( Sun Hwa Ha,Jai Hyun Kim,Young Ki Paik,Jong Bum Kim,Jong Guk Kim,Young Soo Hwang ) 한국응용생명화학회 1998 Applied Biological Chemistry (Appl Biol Chem) Vol.41 No.1

3-Hydroxy-3-methylglutaryl-CoA reductase (HMGR) catalyzes the conversion of HMG-CoA to mevalonic acid, the first intermediate of isoprenoid biosynthetic pathway in plants. The enzyme was solubilized with 0.4% Brij (polyoxyethylene ether) W-1 from a microsomal fraction of etiolated rice seedlings (Oryza sativa L.) in which its maximal activity was observed on the fourth day after germination. HMGR was purified to near homogeneity by employing (NH₄)₂SO₄ fractionation plus chromatographic procedures including DEAE-Sephadex A-50 and HMG-CoA-hexane-agarose affinity column. The size of the purified enzyme was estimated to be 55 kDa when judged by SDS-PAGE analysis with silver staining method. The apparent K_m and V_(max) values for HMG-CoA were determined to be 180 μM and 107 pmol/min/㎎, and those for NADPH were 810 μM and 32.1 pmol/min/㎎, respectively.

고추에서 분리한 Cinnamate 4-Hydroxylase 유전자의 분자생물학적 특성

김계원,하선화,조강진,김은주,이민경,유재주,김종국,이신우,Kim Kye-Won,Ha Sun-Hwa,Cho Kang-Jin,Kim Eun-Ju,Lee Min-Kyung,Yu Jae-Ju,Kim Jong-Guk,Lee Shin-Woo 한국식물생명공학회 2005 식물생명공학회지 Vol.32 No.3

본 연구는 고추열매의 capsaicinoid 생합성 조절 기작을 연구하고자 general phenylpropanoid pathway의 2번째 단계에 작용하는 것으로 알려진 3종류의 c4h cDNA clone을 확보하여 염기서열을 분석한 결과, pc4h1와 pc4h2의 크기는 각각 1,775 bp와 1,655 bp으로 505개의 아미노산으로 구성된 펩티드를 암호하는 full length의 ORF를 갖추고 있었으나 pc4h3는 5'-말단의 coding 영역 일부가 소실 되었다. 이들은 공히 모든 cytochrome P450 효소에서 진화학적으로 보존되어 있는 3종류의 conserved region 즉 domain 1(proline-rich region), domain 2 (threonine-containing binding pocket for the oxygen molecule), 그리고domain 3(heme binding region)을 포함하고 있었으며 evolutionary tree분석을 통하여 pc4h1와 pc4h2는 모두 Class I에 속하는 것으로 서로 간에는 95.8%의 유사성을 나타내어 거의 동일한 유전자인 것으로 조사되었다. 특히 Class II로 분류된 Citrus sinensis C4H1과 Phaseolus vulgaris C4H와는 단지 64.9에서 64.5%의 homology를 나타내었다. 또한 pc4h2는 상처를 주지 않은 조직에서는 비교적 적은 양의 mRNA 발현수준을 보였으나 상처를 가한 후 6시간의 열매에서는 400%, 뿌리에서는 200%까지 그 발현 양이 증가하였다. 이와 유사하게 pc4h1의 전사량도 상처에 의하여 유도는 되나 basal level이 pc4h2보다 높아서 발현증가 비율은 그다지 높지 않았다. 반면에 pc4h3 유전자는 상처를 주지 않은 모든 조직에서 거의 발현되지 않았으며 상처에 의하여서도 전혀 반응을 하지 않았다. Three different cDNAS for cinnamate 4-hydroxylase (C4H) which are involved in the second step of the general phenylpropanoid pathway were isolated and designated as pc4h1 (1,755 bp), pc4h2 (1,655 bp), and pc4h3 (1,316 bp), respectively. The nucleotide sequence analysis revealed that both pc4h1 and pc4h2 clones encode polypeptides of 505 amino acids frame but pc4h3 clone was truncated at the 5'-end of coding region. The alignment of the deduced amino acid sequences showed that PC4H1 and PC4H2 are highly homologous (95.8% identical) with each other and contain three conserved domains which are typical in cytochrome P450 monooxygenase: proline-rich region, threonine-containing binding pocket for the oxygen molecule, and heme binding region. In addition, result of the phylogenic tree analysis revealed that both pepper C4Hs belong to Class 1. pc4h2 transcription was strongly induced in wounded fruit (400%) and root (200%) relative to its very low basal level but not in leaf or stem tissue. In case of pc4h1, the basal level of transcription was higher than pc4h2 but induction by wounding was lower in fruit and root while leaf and stem tissues did not respond to wounding. The basal level of pc4h3 transcripts was not, if any, detectable and response to wounding was not observed.

플라보노이드 대사 조절을 통한 화색 변경

임선형(Sun-Hyung Lim),김재광(Jae Kwang Kim),김동헌(Dong Hern Kim),손성한(Seong-Han Sohn),이종렬(Jong-Yeol Lee),김영미(Young-Mi Kim),하선화(Sun-Hwa Ha) 한국원예학회 2011 원예과학기술지 Vol.29 No.6

화색은 화훼 육종의 주요한 목표형질이다. 최근 유전공학 기술의 발달로 기존의 전통육종에서는 볼 수 없었던 파란장미와 파란카네이션과 같은 새로운 화색 개발이 성공적으로 보고되었다. 플라보노이드 생합성에 관해 축적된 지식기반 연구 결과를 바탕으로 새롭고 독특한 형질의 화색을 도입하는 것이 가능하게 된 것이다. 이러한 화색변경은 플라보노이드 대사경로의 조절, 즉 내재유전자의 발현조절 및 새로운 플라보노이드 합성 또는 특정 플라보노이드 합성을 위한 외래유전자의 추가도입과 플라보노이드 대사 전체를 조절하는 전사인자의 도입을 통해서 이루어져 왔다. 그러나 보다 실증적으로 이러한 플라보노이드 대사를 조절하기 위해서는 작물별 내재 플라보노이드의 조절 기작에 대한 이해를 바탕으로 목표로 하는 플라보노이드 합성을 위해 보다 정교한 대사흐름의 조절이 요구된다. 본 총설에서는 화훼작물의 화색변경 성공 예들을 자세히 소개하고 그 요인 분석을 통해 향후 더 성공적인 화색변경의 전략을 수립하는데 도움이 되고자 한다. Flower color is one of the main target traits in the flower breeding. Recently, technological advances in genetic engineering have been successfully reported the flower colors, such as blue roses and blue carnations that are impossible to develop by traditional breeding. Accumulated knowledge-based approaches for flavonoid biosynthesis enabled to introduce novel and unique colors into flowers. These flower color modifications have been made through the regulation of flavonoid metabolic pathway - control of endogenous gene expression and introduction of foreign genes to produce novel and specific flavonoids - and the introduction of transcription factors that are known to regulate sets of genes being involving in the flavonoid biosynthetic pathway. More empirical regulation of the flavonoids metabolism requires the understanding for regulatory mechanism of intrinsic flavonoids depending on the flower crops and the very sophisticated control of flavonoid metabolic flow. In this review, we summarized successf ul examples of flower color modification. It might be useful to deduce the strategy for the creation of exquisite colors in flower plants.

바이러스 질병 예방을 위한 식물 경구 백신 연구 동향

한범수,박종석,김형국,하선화,조강진,김용환,김종범,Hahn, Bum-Soo,Park, Jong-Sug,Kim, Hyeong-Kuk,Ha, Sun-Hwa,Cho, Kang-Jin,Kim, Yong-Hwan,Kim, Jong-Bum 한국식물생명공학회 2004 식물생명공학회지 Vol.31 No.2

Transgenic plants have been studied as delivery system for edible vaccine against various diseases. Edible plant vaccines have several potential advantages as follows: an inexpensive source of antigen, easy administration, reduced need for medical personnel, economical to mass produce and easy transport, heat-stable vaccine without refrigerator, generation of systemic and mucosal immunity and safe antigen without fetal animal-virus contaminants. The amount of recombinant antigens in transgenic plants ranged from 0.002 to 0.8% in total soluble protein, depending on promoters for the expression of interested genes and plants to be used for transformation. Throughout the last decade, edible plant vaccine made notable progresses that protect from challenges against virus or bacteria. However edible plant vaccines have still problems that could be solved. First, the strong promoter or inducible promoter or strategy of protein targeting could be solved to improve the low expression of antigens in transgenic plants. Second, the transformation technique of target plant should be developed to be able to eat uncooked. Third, marker-free vector could be constructed to be more safety. In this review we describe advances of edible plant vaccines, focusing on the yields depending on plants/promoters employed and the results of animal/clinical trials, and consider further research for the development of a new plant-derived vaccine.

임상용어의 의미적 상호운영성을 위한 매핑 도구

이인근(In Keun Lee),홍성정(Sung Jung Hong),조훈(Hune Cho),김하선(Hwa Sun Kim) 대한전기학회 2011 전기학회논문지 Vol.60 No.1

Most of the terminologies used in medical domain is not intended to be applied directly in clinical setting but is developed to integrate the terms by defining the reference terminology or concept relations between the terms. Therefore, it is needed to develop the subsets of the terminology which classify categories properly for the purpose of use and extract and organize terms with high utility based on the classified categories in order to utilize the clinical terms conveniently as well as efficiently. Moreover, it is also necessary to develop and upgrade the terminology constantly to meet user’s new demand by changing or correcting the system. This study has developed a mapping tool that allows accurate expression and interpretation of clinical terms used for medical records in electronic medical records system and can furthermore secure semantic interoperability among the terms used in the medical information model and generate common terms as well. The system is designed to execute both 1:1 and N:M mapping between the concepts of terms at a time and search for and compare various terms at a time, too. Also, in order to enhance work consistency and work reliability between the task performers, it allows work in parallel and the observation of work processes. Since it is developed with Java, it adds new terms in the form of plug-in to be used. It also reinforce database access security with Remote Method Invocation (RMI). This research still has tasks to be done such as complementing and refining and also establishing management procedures for registered data. However, it will be effectively used to reduce the time and expenses to generate terms in each of the medical institutions and improve the quality of medicine by providing consistent concepts and representative terms for the terminologies used for medical records and inducing proper selection of the terms according to their meaning.