20세기 전반 능주 장범루의 서당 운영

오성 목포대학교 교육대학원 2009 국내석사

Jangbeomru is a Noojeong(pavilion) located in Ohryu-ri, Leeyang- myeon, Hwasoon-gun, south Jeolla province, which commands a fine view. That is one of local cultural assets in Hwasoon-gun. In 1873, Min Chi-gyu built it in commemoration of Min Jeong-sa, his ancestor, and Min Jeong-ho who was his descendant named it Jangbeomru after Jang Gong-ye and Beom Hee-moon. The nine generations of the former's family lived a peaceful life, and the latter was loyal to his brethren. Unlike other pavilions, there was a Jegak in Jangbeomru, which is called Youngmojae. The descendants had a memorial service for Sim Eui-jae at a fixed date. In addition to cherishing the memory of Sim Eui-jae, Jangbeomru functioned as an educational place. That was built primarily to have a memorial service for Sim Eui-jae, but that served as one of informal local educational institutions, namely a village schoolhouse called Seodang. Jangbeomru Seodang provided traditional Confucian education. A family had continued to hold the position of the schoolmaster over about 10 decades since the village schoolhouse opened. Students who lived in remote areas as well as adjacent regions received education there, and the village school was open in particular seasons due to the regional characteristics. That was open from November to April of the lunar calendar when the farming season began, and that was closed temporarily to celebrate the New Year and the 15th of January. Like typical village schoolhouses, Jangbeomru Seodang offered systematic Sino-Korean education and made a great contribution to local development by producing a lot of competent people. As a pavilion, Jangbeomru was in pursuit of the unity of the race, and that rendered great services to nurturing scholars and local development as a Seodang. But it is a shame that Jangbeomru is disregarded at present, and few systematic studies have ever focused on it. In the future, a lot of research efforts should be directed into that. 장범루(張范樓)는 전남 화순군 이양면 오류리의 경치가 수려한 곳에 자리를 잡은 누정으로서 화순군 향토문화재로 지정되어 있다. 1873년 민치규(閔致圭)가 선조 민정사(閔挺泗)를 기리기 위해 누정을 지었고, 후손 민정호가 구세(九世)가 화목한 장공예의 장과 동족들에게 의리를 지킨 범희문의 범을 따서 장범루라고 명명하였다. 장범루는 다른 누정들과는 다르게 누정 안에 영모재라는 제각이 있다. 후손들은 심의재를 추모하기 위해 날짜를 정하여 영모재에서 제를 올리고 있다. 장범루는 심의재를 추모하는 기능 외에도 누정의 교육적 역할을 행하여 서당을 운영하였다. 이것 또한 건립할 때의 취지에 따라 심의재를 추모하는 것으로 나중에는 지역사회의 비공식 교육기관이였음을 보여준다. 장범루의 서당은 전통유교적인 사상에 입각하여 교육하였다. 서당에서 교육을 했던 훈장은 서당교육이 시작한 이래로 약 100년 동안 한 집안에서 대대로 세습되었다. 학생들은 근처 마을 뿐만 아니라 먼 곳으로부터 와서 서당교육을 받았다. 서당은 지역적 특성에 맞게 계절적으로 운영되었다. 음력 11월에 시작하여 중간에 설과 정월대보름에 잠깐 휴식하였다가 농사철이 시작되는 음력 4월까지 교육을 하였다. 장범루 서당은 전통 서당과 같이 체계적인 한문교육을 하였다. 그런 결과 서당교육만으로 지역인재를 많이 배출하여서 지역발전에 크게 이바지하였다. 이러한 사실을 통해서 장범루는 누정으로 동족인들의 화합을 추구하였고, 서당으로서 후학양성과 지역발전에 크게 기여하였다는 것을 알 수 있다. 그러나 다만 한 가지 아쉬운 점은 지금의 장범루가 소홀히 대접받고 있고 제대로 된 연구가 없는 실정이다. 앞으로 이 분야에 대해 더 많은 연구가 이루어져야 할 것이다.

朝鮮後期 商人硏究

오성 西江大學校 大學院 1988 국내박사

비정질 실리콘 박막의 고온 고상 결정화

오성재 (吳晟在) 弘益大學校 大學院 2006 국내석사

최근 능동 영역 액정 표시소자(Active Matrix Liquid Crystal Displays : AMLCD)와 능동 유기 전계 발광 소자(Active Matrix Organic Light Emitting Diode, AMOLED)에 적용되는 다결정 Si 박막 트랜지스터(Poly-Si Thin Film Transistors)에 대해 많은 연구가 진행되고 있다. 현재 상용화되는 대부분의 TFT는 비정질 Si 박막 트랜지스터를 이용하고 있다. 비정질 Si 은 제작이 용이하고 고온 공정에 필요치 않기 때문에 값싼 유리 기판을 사용할 수 있다는 장점을 가지고 있다. 그러나, 비정질 Si은 낮은 전자 이동도(≤0.5cm2/Vs) 및 구동회로와 스위칭 소자를 각각 구현해야 하는 단점을 가지고 있으며 고집적화, 대면적화 하는 추세에 적용이 어려운 문제점을 안고 있다. 그러므로 비정질 Si 박막 트랜지스터에 비해 전계 효과 이동도가 우수하고 구동회로와의 집적이 가능한 장점을 지닌 다결정 Si 박막 트랜지스터가 필요하게 된다. 또한, 다결정 박막 트랜지스터의 제작 시 기판으로서 유리의 적용을 위해 저온 공정(<450oC)을 필요로 한다. 지금까지 다양한 저온 poly-Si TFTs의 제조 공정이 연구되고 있으나 엑시머 레이저 결정화(Excimer Laser Crystallization : ELC) 방법이 가장 효과적인 공정으로 알려져 있다. 그러나 레이저 빔 자체의 조사량이 불균일하다는 레이저 시스템 상의 문제점과, 조대한 결정립을 얻기 위한 레이저 에너지 밀도의 공정 영역이 극히 제한되어 있다는 레이저 공정상의 문제점, 그리고 대 면적에 shot자국이 남는다는 문제점을 가지고 있다. 따라서 본 연구에서는 Non-laser 방식의 고상 결정화방법을 통하여 결정화 시 핵 생성(nucleation) 및 성장(growth)에 관한 kinetics 및 결정 격자결함의 회복 및 제거에 관하여 중점을 두었다. 먼저 Si wafer 위에 PECVD로 형성된 비정질 Si 박막을 600~1000oC의 결정화 온도에서 고온 고상 결정화를 실시하였으며, 핵 생성과 성장에 관한 기본적인 kinetics 및 결정 격자결함의 변화를 온도와 시간에 따라서 확인할 수가 있었다. 결정화 온도와 시간이 증가함에 따라 결정 격자결함은 급격히 제거되었으며, 핵 생성과 성장의 시간이 감소하였다. 또한 동일 결정화 시간에서 온도가 증가함에 따라 핵 생성 구간, 결정화가 완료되는 구간, 결정 격자결함이 제거되는 구간으로 구분이 되었으며 핵 생성 및 성장의 에너지보다는 결정 격자결함의 제거를 위한 에너지가 더 많이 요구되는 것을 알 수가 있었다. 본 연구를 통하여 현재 Mo, steel 등의 기판을 사용하여 결정화 온도에 영향을 받지 않는 Flexible Display의 개발과 더불어 AMOLED의 대면적화와 균일도를 가지는 고품위 다결정 Si TFTs의 제조에 기여를 할 것으로 판단되고, 고상 결정화에 대한 전반적인 kinetics를 확인할 수가 있었다. For application to active matrix liquid crystal displays (AMLCDs) and active matrix organic lighting emitting diode (AMOLED), a low temperature process for the production of poly-Si is required to permit the use of glass substrates. Polysilicon films best suited for TFTs are made by crystallization of a-Si:H precursor films. Crystallization techniques include furnace annealing, rapid thermal annealing by lamp heating, and laser crystallization. Furnace annealing produces highly uniform polysilicon films over large areas, and is a proven batch process. Because the strain points of affordable substrate glasses lie about 600oC, crystallization and further processing are restricted to temperatures at or below~600oC, which requires crystallization and ion-implant annealing times as long as 20hrs. Catalyzed crystallization can reduce this time to~5hrs, which still is long when compared to the process step throughput of one plate per minute desired of the single-substrate cluster tools employed in the manufacture of active-matrix liquid-crystal displays. To find a fast and furnace-based crystallization process for large areas of low-cost substrates to enable the integration of driver circuits for active matrices has been the primary goal of our research. In this thesis work crystallization kinetics including nucleation and growth and the microstrucutres obtained at higher temperatures were investigated. SPC experiments were conducted using a tube furnace at temperatures ranging from 600oC to 1,000oC. The samples used consist of layered structures of 50nm-thick a-Si/500nm-thick SiO2/Si wafer. Crystallization kinetics becomes dramatically rapid as the crystallization temperatures increases to higher temperatures. According to TEM observation the grain size decreases as the annealing temperature increase up to 800oc. It is, however, not so sensitive to the annealing temperature beyond 800oC. Since crystallization kinetics becomes very rapid and since the heating rate of a conventional furnace is slow phase transition to polycrystalline silicon may occur in the course of heating-up, and thus the desired microstructure formed at high temperatures cannot be reflected.

중국어권 한국어 학습자를 위한 관용표현 교육 방안 연구

오성아 충북대학교 2017 국내박사

The importance and understanding of foreign languages are escalating in this age of globalization. Particularly in China, after the establishment of diplomatic ties with Korea, an interest in the Korean language is gradually increasing. Accordingly, more Chinese people have been learning the Korean language, making them the largest group of Korean language learners amongst all foreign nationalities. The main reason why so many Chinese people learn Korean is that they might think Korean is easy to learn because of its Chinese origins. Due to the high proportion of Chinese characters in the Korean lexicon (hereby known as hanja), Chinese native speakers can learn Korean with more ease than speakers of other languages. Even though it is not an absolute guarantee to mastering the Korean language for many reasons, it is true that Korean is much easier to learn for Chinese native speakers than speakers of other languages. However, there are still stark differences between hanja and the modern Chinese language. Even though the characters look the same, some hanja and Chinese characters have slightly different meanings and others have completely different meanings. Thus, Korean language learners may be confused. One of the confusing factors is “idiomatic expressions”. Idiomatic expressions are often prahses or sayings habitually used by native speakers that convey the values, ways of thinking, feelings, and history of the nation. Without fully understanding these factors, foreigners could understand idiomatic expressions only by its surface meaning. The idiomatic expressions are not only used frequently in daily life but also appear on the TOPIK. Unsurprisingly, idiomatic expressions account for a great part in learning Korean. But, at the beginner’s level, the need for idiomatic expressions in communication is not as necessary for beginners than it is to higher Korean level learners. Korean idiomatic expressions are one of the difficult parts in learning Korean because they are infused with Korean cultural, historical and social meanings. If learners can properly use and express these expressions, it shows their strong competency in the Korean language. But learners cannot and do not have to learn all idiomatic expressions as beginners. So, as learners progress in their Korean level, they shift from learning everyday expressions to expressions embedded with deeper cultural and historical meanings. The word list in this thesis is organized by the idiomatic expressions’ frequency of usage and level of difficulty. This thesis aims to find teaching methods that teachers who teach Korean specifically to foreigners can use to let Chinese learners understand a large number of Korean idiomatic expressions in systematic, concrete, and effective ways. In doing so, this paper focuses on constructing a proper syllabus and methods according to the learners’ respective levels. This thesis analyzes the idiomatic expressions in Korean texts used not only in Korea but also in China, and the learners in both countries are surveyed. Based on the results, this thesis proposes an effective teaching syllabus for Chinese learners of Korean. In addition, the paper recommends practical teaching methods and plans so learners can learn idiomatic expressions in different ways. In further study, when making the syllabus, the research considers the learners’ awareness of idiomatic expressions. Also, Korean idiomatic expressions are analyzed alongside with their Chinese counterparts in order to understand their similarities and differences.

통일교의 영성교육에 대한 연구 : 통일교의 심정교육을 중심으로

오성 선문대학교 신학전문대학원 2007 국내석사

본 연구는 21세기를 들어서면서 현대사회가 요구하는 바가 과학의 발달에 따른 물질적인 풍요를 넘어서서 정신적인 가치를 추구하고 있음을 절실히 느끼면서, 오늘의 종교가 인간의 영성계발과 교육을 위해 어떠한 인식 가운데서 어떠한 특징을 지니고 있으며, 이에 통일교의 영성과 영성교육은 어떠한 내용과 특징을 갖고 있는지를 알아보고자 하는 데 그 목적이 있다. 기독교에서 말하는 영성을 다른 한마디로 표현한다면 ‘신성(神性)’이라고 말할 수 있다. 그리고 하나님의 신성이 그대로 전개되어 창조된 것이 피조만물이요 인간이라고 볼 때, 영성의 형성은 신의 성품을 형성하는 것으로 해석될 수 있다. 즉 ‘신성을 사람 안에 이루는 것이다’라고 표현할 수 있다. 통일교의 영성에 대한 이해는 통일원리의 신성에 대한 이해로부터 시작된다. 통일원리에서 말하고 있는 신성은 심정(心情), 로고스(이법), 창조성(創造性) 등 크게 세 가지로 이야기하고 있다. 그러므로 통일교의 영성의 특징은 체휼일체성(體恤一體性), 이법성(理法性), 창조성(創造性)을 들 수 있다. 이어 기독교의 영성에 대한 일반적인 개념과 시대적인 변화과정 및 현대적 개념이해에 대해 다루었다. 또한 현대적 영성의 특징을 통전성, 프락시스성, 해방성, 일상생활성 등 네 가지로 특징지워 설명하였다. 다음으로 통일교의 영성교육은 하나님의 심정을 중심한 개인의 인격완성을 위한 교육으로 이를 심정교육이라고 말한다. 그러므로 통일교의 영성교육은 하나님의 심정에 대한 이해가 우선되어야 한다. 통일교의 영성교육의 특징은 첫째, 영성이 ‘하나님과 인간 사이에서 형성되는 관계성을 총칭한다’고 할 때 하나님과 인간의 관계의 근본인 부자의 관계를 가르치는 교육이다. 둘째, 통일교의 영성교육은 하나님의 심정을 중심한 인격완성을 위한 전인교육이다. 셋째, 통일교의 영성교육은 상대를 위하여 투입하는 참사랑의 교육이다. 넷째, 통일교의 영성교육은 로고스(말씀)을 중심한 이법(理法)교육이다. 다섯째, 통일교의 영성교육은 말씀을 수육하며 참사랑을 실천해 나가는 실천교육이다. 여섯째, 통일교의 영성교육은 끝없는 참사랑의 투입과 사랑의 주관을 통한 창조교육이다. 일곱째, 통일교의 영성교육은 가정적 사위기대를 중심으로 3대 상사랑과 4대 심정권을 완성해 나가는 참가정교육이다. 여덟째, 통일교의 영성교육은 창조이상가정을 중심한 천국생활교육이다. 아홉째, 통일교의 영성교육은 영인체의 완성과 더불어 영계에서의 영원한 삶을 위한 교육이다. 이러한 특성을 갖고 있는 통일교의 영성교육은 심정적 체휼의 과정을 통해 이뤄지는데 체휼의 과정은 마음의 영점기준, 암시, 몽시, 계시, 묵시, 영적인 역사, 육신의 순화 등으로 발전해 나간다. 그리고 통일교의 영성교육의 한 실제로 문형진의 ‘천일국백성수련’을 그 예로 들었다. 다음으로 기독교의 영성교육에 대해서는 먼저 일반적인 영성과 교육의 문제 및 기독교의 현대적 영성교육의 특징을 분석하고 그 실제적인 한 모형으로 마리아 해리스(Maria Harris)의 영성의 춤(Dance of the Spirit : the Seven Steps of Women’s Spirituality)을 제시하였다. 우리는 위와 같은 영성의 이해와 영성교육을 통하여 하나님을 닮아난 하나님의 참자녀로 거듭날 수 있게 되고, 이 영성교육이 인간이 가야 할 참된 길을 제시해 주는 필연적인 교육임을 알게 된다. With the start of the 21st century, we have come to see that modern society is pursuing spiritual values beyond the material wealth that comes with the advancement of science. On this background, this paper will examine the perceptions adopted by contemporary religions in developing human spirituality and the characteristics of spirituality education by such religions, and analyze the contents and characteristics of the Unification Church’s perception on spirituality and its spirituality education. In Christianity, spirituality is expressed alternatively as "divinity." Christians believe that God’s divinity was developed and made manifest in creation and human beings. Thus, the formation of spirituality can be understood as the formation of God’s character. In other words, divinity is formed within man. The Unification Church’s understanding of spirituality begins from the understanding of God’s divinity as defined in the Unification principles. The divinity explained here is composed of three elements: heart, logos (natural law) and creativity. Consequently, some characteristics of spirituality in the Unification Church are the ability to incorporate or embody God’s heart, rationality and creativity. This paper examined the general concept of Christian spirituality, the changes it went throughout the ages and the current understanding of spirituality in Christian society. Spirituality today is characterized by integrity, praxis, liberation, and everydayness. The purpose of spirituality education in the Unification Church, or ‘education of heart,’ is to perfect an individual’s character centering on God’s heart. Consequently, the spirituality education in the Unification Church puts priority in understanding God’s heart. Spirituality education in the Unification Church defines spirituality as the general term for the relationship formed between God and man, and teaches that the relationship between God and man is fundamentally that of one between a parent and child. Second, it engages with the entire being of a person enabling individual’s to perfect their character centering on God’s heart. Third, it is an education based on true love, of investing oneself for one’s partner. Fourth, it is rooted in rationality centering on God’s Logos (Word). Fifth, it is a practical education for embodying the Word and practicing true love. Sixth, it teaches of infinitely investing true love and governing creation in love. Seventh, it is an focused on creating a true family which perfects the three object purposes and the four great realms of heart centering on the four-position-foundation in a family. Eighth, it teaches how to lead one’s life in the kingdom of heaven centering on the ideal family that was to be established at the time of creation. The purpose of spirituality education in the Unification Church is for human beings to perfect their spirit self and to learn how to lead an eternal life in the spirit world. Spirituality education in the Unification Church occurs through experiential learning of the heart. This process develops in stages from the zero-point of the mind, to hints, dreams, revelations, divine revelations, spiritual works and ultimately the purification of the physical body. One example of this is the "Cheon Il Guk citizen’s workshop" organized by Hyung-jin Moon. In examining the spirituality education in Christianity, this paper examined the problems in spirituality in general and in the education of spirituality. Then, it analyzed the characteristics of modern Christian spirituality education and introduced Maria Harris’ Dance of the Spirit (the Seven Steps of Women’s Spirituality) as one model being employed in Christianity. Through an understanding of the afore-mentioned spirituality and spirituality education, we can be reborn as God’s children that resemble Him, and understand that the spirituality education of the Unification Church essentially points to the true way which should be followed by all human beings.

自家骨移植에 依한 齒槽骨下 盲囊處置의 實驗的 硏究

오성용 서울大學校 大學院 1969 국내박사

4,4',4''-Tris[1-naphthyl(phenyl)amino]-triphenylamine 증착박막의 분자배향 및 유기발광소자의 성능향상

오성 부산대학교 2007 국내석사

This study is for the flatness and the improvement of the molecular ordering on the 1-TNATA thin film under the electromagnetic field and the annealing effects. AFM analysis is observed the surface morphology and Raman spectra showed the molecular ordering. We intended to make higher performance OLED using the treated 1-TNATA thin film.1-TNATA was deposited onto ITO glass substrates via a vacuum process. As it is recognized that highly oriented organic/metal-organic films are of great importance for device purposes, it is essential to have high quality films for device purposes. AFM and Raman spectra analysis were used to characterize the topology and structure-oriented molecular ordering. 1-TNTA films. A source meter was used to observe the current-voltage characteristics of the ITO/1-TNATA/Al device. While 1-TNATA thin films deposited under electromagnetic field were not structurally oriented according to the Raman spectra results, 1-TNATA thin films after post-deposition annealing at 110℃ were oriented. According to the AFM images, 1-TNATA thin films after post-deposition annealing at 110℃ had a smoother surface than those deposited without electromagnetic field. Current-voltage performance of 1-TNATA thin films was improved due to the enhanced electron mobility in the well-aligned 1-TNATA films. Especially surface roughness is very important to enhance the stability and efficieny of OLED. As a result, we observed the better I-V curves caused by the better molecular orientation and flatness in electromagnetic field treated 1-TNATA. Furthermore, we found the influences of the luminescent efficiency, the turn-on voltage and brightness in OLED subjected to the 1-TNATA thin films which were fabricated using various temperatures (90,110,130℃) and magnetic filed (~6mT). No change with the range of the emitting area is observed.

Phylogenetic analysis of begomovirus and associated betasatellite occurred in Korea and characterization of their interactions and pathogenicity : 국내에 발생하는 베고모바이러스와 베타 DNA의 계통적 분석 및 감염성 클론을 이용한 상호작용과 병원성 규명

오성 배재대학교 대학원 2017 국내박사

Previously, Tomato yellow leaf curl virus (TYLCV)-IL[KR:Bus] (accession number GQ141873), -IL[KR:Bos] (accession number GU325634), -IL[KR:Hwas] (accession number GU126513), and Honeysuckle yellow vein virus (HYVV)-[KR:DJ] were isolated, respectively, from tomato (Solanum lycopersicum) plants in Korea. To construct TYLCV infectious clones, full-length of respective TYLCV genomes were amplified by polymerase chain reaction (PCR) and cloned into binary vector pRI101-AN, respectively. Using the 1mer clone, 2mer genomes of each isolate were constructed by restriction enzyme digestion and ligation. The resulting constructs were designated pRI-TYLCV-Bus, pRI-TYLCV-Bos and pRI-TYLCV-Hwas. In the case of HYVV, PCR amplified unit-lengths of DNA-A genome of HYVV-[KR:DJ] were cloned into binary vector pRI101-AN. Using the 1mer clone, 1.3mer and 2mer genomes were constructed by restriction enzyme digestion and ligation. The resulting constructs were designated pRI-HYVV-DJ. To prove the infectivity of each clone, respective constructs were transformed into Agrobacterium tumefaciens cells and the selected transformants were agro-inoculated into young leaves of clonescontaining dimer construct of respective isolates caused pronounced disease symptoms such as plant stunting, downward leaf curling and crinklingn.InHYVV,1clones caused pronounced disease symptoms in To discriminate the clone’s pathogenicity quantitatively, SYBR Green-based real-time PCR was performed for viral quantification using V1 gene DNA content in agro-inoculated leaves that were collected at weekly intervals for 4 weeks. Regression analysis obtained the standard curves by plotting Ct values over the logarithm of the amount of V1 DNAs present in a dilution series of plasmid containing the full-length HYVV-[DJ] genome. Equation of the HYVV-V1 DNA standard curve was used to quantify V1 gene DNA concentration in agro-inoculated plants with each clone. A real-time PCR assay showed that the accumulation of V1 DNAs in HYVV-[DJ]-1.3mer inoculated plants reached the peak level at 4 weeks post-inoculation. The amount of V1 DNA was significantly more than that in HYVV-[DJ]-2mer inoculated plants. Nevertheless, the accumulation of V1 DNAs reached the peak level at 3 weeks post-inoculation and then decreased slowly from 4 weeks post-inoculation. Considering the real-time PCR results, a significant difference was found in the accumulation virus and its DNA components between the analyzed plants inoculated with each clone, indicating that the difference in clones’ pathogenicity correlates with their virulence. A polymerase chain reaction (PCR) using two sets of primers designed from published Tomato yellow leaf curl virus (TYLCV) genomes was developed to distinguish from the TYLCV-IL groups. The specificity of the two sets of primers was proven by testing against control TYLCV genomes and the symptomatic leaves of 34 different tomato cultivars naturally infected with TYLCV in greenhouses. One set for amplified full-length genome fragments from all the 34 tomato cultivars. Another set for TYLCV-ILgroup-II strain specific primers amplified target DNA fragments from only 9 tomato cultivars. Digestion by BglII and EcoRV of the PCR amplicons produced restriction fragment length polymorphism (RFLP) pattern the TYLCV-IL group-I with two fragments from the TYLCV-IL group-II with no digested fragment. PCR coupled with BglII and EcoRV digestion confirmed that the 9 tomato cultivars were infected with the TYLCV-IL group-II and the remained 25 tomato cultivars were infected with the TYLCV-IL group-I. Since the begomovirus C4 protein is known to associate with symptom severity (SS) and effective systemic infection, it is interesting to measure C4 protein level in tomato (Solanum lycopersicum) cultivars infected with Tomato yellow leaf curl virus (TYLCV). In this study, the full-length of the C4 protein gene of the TYLCV and the Honeysuckle yellow vein virus (HYVV), respectively, were expressed in Escherichia coli. To determine the antigenic characteristics of recombinant C4 (rC4) protein, polyclonal rat antibody was produced against the purified rC4 protein with His tag as an immunogen. In terms of the specificity, TYLCV-rC4 antiserum reacted well not only with TYLCV-rC4 but also with HYVV-rC4 antigens in Western blot, and vice versa. To determine the infection rate qualitatively and quantitatively, PCR and real-time qPCR assays were performed with eight tomato cultivars agro-inoculated with a TYLCV infectious clone and a naturally infected tomato cultivar with TYLCV. The real-time qPCR results revealed that the accumulation level of viral DNA was much higher in plants of susceptible cultivars than in plants of tolerant cultivars. It indicated that there is a negative correlation between the virus DNA accumulation and the presence of Ty resistance genes in the tolerant cultivars. Nevertheless, higher virus DNA accumulation was not necessarily matched with more severe symptoms observed in the susceptible tomato cultivars. To quantify the sensitivity of the TYLCV-rC4 antibody, enzyme linked immunosorbent assay (ELISA) was performed with four selected plants of each cultivar showing TYLCV positive by PCR. ELISA results revealed that the C4 protein accumulation level was much higher in plants of the susceptible cultivars than in plants of the tolerant cultivars. In general, more C4 protein accumulated in susceptible cultivars showing higher SS rating, suggesting that the amount of C4 protein in plants positively related with symptom severity of the cultivar. The complete nucleotide sequences of Honeysuckle yellow vein virus (HYVV-HB) genome DNA and a betasatellite DNA (HYVB-HB) were determined from honeysuckle (Lonicera japonica) leaves showing characteristic yellow vein mosaic symptoms in the Hanbat Arboretum, Daejeon, Korea. HYVB-HB had high nucleotide sequence identity with Honeysuckle yellow vein mosaic virus (HYVMV)-associated DNA β found in UK (accession number AJ543430: 98.4%), New Zealand (accession number GQ131809: 98.2), Korea (accession number JQ728545: 98.2) and Japan (accession number AB236427: 97.3). Previously, we identified Tomato yellow leaf curl virus (TYLCV)-[Bus] and Honeysuckle yellow vein virus (HYVV)-[DJ] from tomato plants, each found not to be associated with a satellite DNA. To prove pathogenicity and interactions between a begomovirus and noncognate DNA β, we demonstrate co-agroinoculation into Nicotiana benthamiana, using a mixture of infectious clones of TYLCV + DNA β-HB and a mixture of HYVV + DNA β-HB. Genetic recombination is an important event during the evolution of begomovirus species. To determine the possibility of recombinant viruses, we investigated patterns of recombination that occur in 5 weeks long experimental infections of Nicotiana benthamiana plants using two infectious clones containing hybrid constructs (T+H hybrid and H+T hybrid) between TYLCV and HYVV genomes. Like original dimer clones of TYLCV and HYVV genomes, the infectious clones containing T+H hybrid and H+T hybrid constructs developed plant stunting and leaf curling with crinkling when agro-inoculated into N. benthamiana in the infectivity assay. PCR amplification and real-time qPCR analysis supported the infectivity of clones containing T+H hybrid and H+T hybrid constructs. The PCR amplified DNA products from plants agro-inoculated with T+H hybrid construct and H+T hybrid construct (Fig. 6D), respectively, cloned into pRI101-AN vector and determined their nt sequences. We found two recombinants named TH virus from the T+H hybrid construct agro-inoculated plants and HT virus from the H+T hybrid construct agro-inocualted plants. The full-length genome (2774 bases) of HT virus showed 74.1% and 97.3% nt identities, while its IR (300 bases) revealed 79.2% and 75.0% nt identities with those of HYVV-[KR:DJ] and and TYLCV-[KR:Bus], respectively. The full-length genome (2764 bases) of TH virus showed 97.0% and 73.9% nt identities, while its IR (296 bases) revealed 75.5% and 78.5% nt identities with those of HYVV-[KR:DJ] and TYLCV-[KR:Bus], respectively. In between TH and HT viruses their nt sequence identities was 71.4% in the full-length genome and 54.9% in the IR, which are same nt identities of the full-length genome and IR between HYVV-[KR:DJ] and TYLCV-[KR:Bus]. Based on nt sequence analysis, we found that recombination events mainly occurred in IR but not in ORF-coding regions, Based on nt sequence analysis, we found that recombination events mainly occurred in IR but not in ORF-coding. Although 54.9-79.2% nt identities were found in the IR, a stem-loop structure containing nonanucleotide region shows highly conserved nt sequences in recombinat viruses. All the results together indicated that the recombinant TH virus is closely related to HYVV and the recombinant HT virus is closely related to TYLCV. Severe tomato yellow leaf curl disease occurred in a TYLCV-tolerant cultivar (Dafnis) containing Ty- 1 and Ty-3 genes. Infected plants were collected from Buyeo city in Chungnam province, Korea. The complete nucleotide (nt) sequences were determined as 2774 bases, indicative of a monopartite begomoviral genome. A comparison of the ful-length genome with known TYLCV isolates reported in Korea, showed that the virus shared the highest nt sequence identity (98.5%) with TYLCV-IL[KR:Bus] considered as TYLCV-IL group-II. Based on the guidelines of the ICTV the virus has been designated TYLCV-IL[KR:Buyeo]. In addition, a betasatellite DNA was amplified by PCR and cloned, and the nt sequences were determined. Analysis of the complete nt sequences of 1347 bases indicated that the betasatellite (HYVB-tomato) shared 99.9% identity with its closest relative honeysuckle yellow vein betasatellite (HYVB-HB; accession number KC788280) which was identified from honeysuckle plants infected with Honeysuckle yellow vein virus (HYVV).

Molecular characterization of infectious clones of Honeysuckle yellow vein virus (HYVV) and Tomato yellow leaf curl virus (TYLCV) and expression of HYVV and TYLCV genes in Escherichia coli and Nicotiana benthamiana

오성 배재대학교 대학원 2012 국내석사

Two newly emerged begomoviruses were isolated from naturally infected tomato (Solanum lycopersicum) plants grown in greenhouses at Jeju Island and Dangjin in Korea and their genomes were characterized. These viruses-infected plants had very small leaves that curled upward, yellow margins and a leathery appearance, and a bushy and stunted appearance with short internodes. Nucleotide (nt) sequence analysis of their genomes showed that they have a DNA-A component of monopartite begomovirus. Their genomes comprised 2763 and 2764 nucleotides with six open reading frames. The results of nt sequence similarity analysis of DNA-A genome between the two Korean isolates and isolates of Tobacco leaf curl Japan virus (TbLCJV), Honeysuckle yellow vein virus (HYVV), Honeysuckle yellow vein mosaic virus (HYVMV), and Eupatorium yellow vein virus in Japan (EpYVV) showed that they are likely similar to HYVV-[Masuda] (89.4-92.8% nt identity). Consequently, we tentatively propose the two isolates’ names as HYVV-Jeju and -DJ according to the ICTV geminivirus rules. Phylogenetic relationship analysis of 33 DNA-A genome sequences using PAUP* 4.0b10 and MrBayes revealed that HYVV-Jeju and -DJ belong to the Far East Asian begomovirus species complex. Within the Far East Asian begomovirus species complex, HYVV-Jeju and -DJ are distantly related to EpYVV, HYVMV, and TbLCJV groups. Based on the presence of a recombination fragment spanning the C3 ORF, a recombinant origin was suggested for both HYVV-Jeju and –DJ, with parents close to Japanese isolates HYVMV-[SP1:00] and Eupatorium yellow vein virus (EpYVV)-[Suya]. In addition, the presence of a further recombination fragment spanning the IR suggested the parents of HYVV-DJ were close to HYVV-Jeju and EpYVV-[Suya]. Honeysuckle yellow vein virus (HYVV)-[DJ] (HQ189431) isolated from tomato plants, has a DNA-A component of monopartite begomovirus without β satellite molecules. Unit-lengths of DNA-A amplified by PCR, which were synthesized 1-mer (monomer), 1.3-mer and 2-mer (dimer) genomes using restriction enzyme digestion and ligation. The resulting constructs, respectively, ligated into binary vector pRI101-AN. Additionally, the construct containing 2mer cloned into binary vector pCAMBIA1304. To investigate the infectivity of clones, each construct transformed into Agrobacterium tumefaciens cells, which was agroinoculated into young leaves of Nicotiana benthamiana, N. tabacum Xanthi and Solanum lycopersicum. Symptoms appeared as mild or severe leaf curling with plant stunting in these plants except N. tabacum Xanthi. Polymerase chain reaction (PCR) verified a systemic infection of these plants by the agro-infectious clone. Infected leaf tissues of N. benthamiana were cultured as a callus in solid media and used for in vitro HYVV propagation. Two passages of callus culture resulted in maintenance of virus population and proved systemic infection of HYVV. Mutation easily occurs in begomovirus genome. Because of quasispecies-like nature of the Tomato yellow leaf curl virus (TYLCV) populations, it is doubtful that variable populations of TYLCV can be generated in a short period of time for the adaptation in response to changing host plant. To investigate this question, infectious clones of (TYLCV)-[Bus] (GQ141873) isolated from tomato plants were generated to inoculate via Agrobacterium into Nicotiana benthamiana plants. The unit-length of TYLCV-[Bus] genome DNA was amplified by PCR, which was cloned into pRI101-AN or pCAMBIA1304 vector. This cloning resulted in the recombinant vector named pRI-TYLCV-1mer, -1.3mer and -2mer and pCAM-TYLCV-2mer. Each clone was transformed into Agrobacterium tumefaciens cells, which were infiltrated into young leaves of N. benthamiana. In the case of pRI-TYLCV-1mer, -1.3mer and -2mer, all clones did not produce distinctive foliar symptoms on the inoculated plants, except mild plant stunting, in four weeks post-inoculation. In the case of pCAM-TYLCV-2mer, the clone produced mild foliar symptom and mild plant stunting in four week post-inoculation. Regardless of the symptom severity, all inoculated plants were verified to confirm the presence of TYLCV by PCR. TYLCV-V1 gene was amplified from the agro-infected plants by PCR and sequenced its nucleotides. However, there was no difference in nucleotide sequences compared with wild type TYLCV-[Bus]-V1 gene. It suggests that the assumption is not correct under the experimental condition in this study. Recently, two begomoviruses, Tomato yellow leaf curl virus (TYLCV) and Honeysuckle yellow vein virus (HYVV), have occurred in tomato fields in Korea. To determine the antigenic characteristics of begomovirus C4 protein which is known to play diverse functions in virus-host interaction, the entire C4 protein genes of HYVV and TYLCV were amplified by PCR and engineered to be expressed by a bacterial expression vector pET-28a. SDS-PAGE gel revealed that HYVV-C4 and TYLCV-C4 proteins were expressed, respectively, as about 16.4 kDa and 15.9 kDa bands in Escherichia coli, and found mainly in the pellet of the bacterial lysates. The recombinant protein C4 (rC4) fused with His-tag was purified from E. coli using Ni-NTA resin under denaturing conditions. Polyclonal rat antiserum was produced against each purified rC4 protein to use as an immunogen. HYVV-rC4 antiserum reacted well not only with HYVV-rC4 antigen in Western blot but also with TYLCV-rC4 antigen, and vice versa. Each antiserum was further purified to isolate respective immunoglobulin G (IgG). Each IgG had a better resolution than the crude antiserum when tested with rC4. 본 연구는 2008년도에 국내에서 처음 발견된 후, 전국적으로 대유행하며 시설하우스 토마토 농가에 막대한 피해를 입히는 Geminiviridae의 Begomovirus에 대한 저항성 품종 선별과 정확하고 민감성의 진단시스템 구축을 위한 3개의 연구주제를 수행하였다. 첫 번째는, 당진에서 분리된 Honeysuckle yellow vein virus (HYVV-DJ)의 분자생물학적 특성을 규명하였다. Begomovirus의 유사 증세를 보이는 토마토를 당진의 농가에서 채집하였고, V1(Coat protein) gene을 중합효소연쇄반응을 통하여 증폭하였다. 염기서열을 통해 begomovirus에 속한 HYVV인 것을 밝혀냈다. 중합효소연쇄반응을 통해 전체 유전자를 증폭한 후, 저장 vector인 pGEM T-easy vector에 클로닝하였다. HYVV의 6개의 ORF와 1개의 IR의 염기서열을 모두 파악한 후, 기존에 밝혀져 있던 유사한 Begomovirus들과 염기서열 비교를 하였다. PAUP4.0b10 과 MrBayes 방법을 통한 계통수 결과로 이 바이러스는 Far East Asian begomovirus 종에 속함을 밝혀냈다. 또한 HYVV-DJ는 제주도에서 채집된 HYVV-Jeju와 91.9%의 가장 높은 상동성을 보였고, 일본에서 분리된 HYVV-[Masuda]와 89.4%의 높은 상동성을 보였다. RDP3.44 프로그램을 통해 재조합 상태를 비교하였다. 그 결과, Major parent는 HYVV-Jeju, Minor parent는 EPYVV-[Suya]로 밝혀졌으며, HYVV-Jeju와 Minor parent가 같음을 알게 되었다. 이로 보아 HYVV-DJ는 HYVV-Jeju의 Major parent인 HYVMV-[SP1:00]와 Minor parent인 EPYVV-[Suya], 그리고 HYVV-Jeju로 부터 재조합된 바이러스임을 밝혀냈다. 이는HYVV가 빠른 재조합을 보이며 퍼져 나가고 있음을 확인할 수 있었다. 두 번째는, Begomovirus에 대한 저항성 토마토 품종 선별을 위한 HYVV-DJ, TYLCV-Bus의 전장을 식물발현 vector인 pRI101-AN-vector와 pCAMBIA1304 vector에 클로닝하여 감염성 클론을 만들었다. 바이러스들을 효과적으로 발현시키기 위해 pRI101-AN-vector에 1mer, 1.3mer, dimer를 클로닝하였고, pCAMBIA1304 vector에 dimer를 클로닝하였다. Agro-infiltration 통하여 N. benthamiana, N. tabacum Xanthi그리고 Solanum lycopersicum에 접종하였다. 접종 5주 후, N. benthamiana는 pRI-HYVV-1.3mer , dimer의 경우에는 매우 뚜렷한 오그라드는 증상과 생장의 차이를 보였지만, pRI-HYVV-1mer는 정상 N. benthamiana와 차이를 보이지 않았다. 또한 pRI-TYLCV-1mer, 1.3mer, dimer 모두 별다른 증상을 보이지 않았다. 접종 2주 후, pCAM-HYVV-dimer는 pRI-HYVV-dimer보다는 약하지만, 오그라드는 증상과 성장의 차이를 보였고, pCAM-TYLCV-dimer는 잎맥을 따라 약한 황화 증상만을 보였다. 이 식물들에서 중합효소연쇄반응을 통해 모두 V1 gene이 확인됨에 따라 바이러스가 정상적으로 증식함을 확인되었다. Xanthi는 두 바이러스 모두 병징을 관찰할 수 없었으며, Solanum lycopersicum에서 HYVV의 병징을 관찰하였고, 중합효소 연쇄반응을 통해 상엽에 바이러스가 증식했음을 확인되었다. N. benthamiana와는 달리 Xanthi는 HYVV와 TYLCV에 대한 기주가 아님을 밝혀냈다. 추후 infectious clones을 이용한 저항성 토마토품종 screening에 이용될 수 있음을 확인하였다. 세 번째는, 진단시스템 구축을 위해 HYVV-DJ와 TYLCV-Bus 의 C4 gene을 중합효소연쇄반응을 통하여 증폭하고 이 유전자를 대장균 발현 vector인 pET28a vector에 클로닝하였다. IPTG induction를 통해 HYVV-C4(16.39kDa), TYLCV-C4(15.95kDa) 의 단백질들이 침전물에서 형성되는 insoluble 조건에서 발현됨을 SDS-PAGE로 확인하였다. 이 단백질들을 변성조건에서 Ni-NTA resin을 이용하여 정제하였다. 정제된 단백질을 항원으로 쥐에 주입하여 다클론 항체를 생성한 하였고, 대장균에서 발현된 단백질을 이용하여 Western blot 분석을 통해 정상적으로 다클론 항체가 생성되었음을 확인하였다. 또한 HYVV와 TYLCV의 C4 다클론 항체는 서로 교차 결합하였다. 추후 각 항체를 IgG 정제를 한 결과, 더 감응성이 뛰어난 항체를 얻게 되었다.  

심상의 회화적 표현 연구 : 본인 작품을 중심으로

오성 건국대학교 대학원 2011 국내석사

현대미술의 구조 속에서 우리는 예술의 본질과 이를 표현해내는 다양한 방법론적 사고의 필요성을 인식해야 한다. 때문에 심상의 회화표현에 대한 연구는 다양한 예술의 정보와 함께 가장 개성적인 방법으로 작품에 반영 하여 표현 할 수 있다는 점에서 그 목적을 지닌다. 본 연구자는 작품을 통해 대중과 교류하고 공유하는 것, 그리하여 세상과의 단절 속에서 작가의 내면세계만을 고집하는 것을 우리는 지양해야 할 것이다. 또한 현실 속에서 이루어진 정보의 조합을 통해 자신의 내적 체계가 표현 되지 않는다면 그것은 화가의 심상(心象)적 표현이 아닌, 단순한 기억으로 조합시킨 정보복합체에 불과하다. 때문에 내면의 기억과 감성을 적절하게 융화시켜 자신의 정체성을 깨닫고 표현하는 것이 매우 중요하다. 또한 작품에서의 내면적 형상과 색채를 구체화 하는 것 역시 매우 중요한데, 이는 심상의 회화표현으로 구체화 된 이미지가 대상에 따른 비유적 상징체계를 어떻게 형성하느냐 하는 문제인 것이다. 이것은 개인의 정신적 개념과 조형적 개념을 구체화 시킨 이미지, 또는 내면 갈등 속에서의 개성적 이미지를 보다 구체적인 형상으로 만들어 가는 과정의 한 결과물이라 볼 수 있을 것이다. 사회구조가 더욱 분화되고, 다양해짐에 따라 인간의 일탈을 향한 욕망은 더욱 커지기 마련이나, 제한된 현실은 이를 수용할 만한 포용력을 가지지 못한다. 이 때문에 현실에서의 자아가 가진 환상에 대한 상황적 표현은 제한되어 있어 화면에서의 의식 ․ 무의식의 이미지 연결이 조화롭지 못할 수 있다. 본 연구자는 심상에 있어 비유적 표현방법을 사용해 현실과 환상의 관계를 내면의 의식으로 이미지화시켜 재구성 하였으며, 다양한 상징적 색채의 이미지 요소들에 초점을 맞추어 연구하였다. 또한 가시적으로 보여지는 형상과 내면의 심리적인 추상이 서로 이원화되지 않고 상호작용을 통해, 상대적 융합을 통한 미적가치를 유추할 수 있게 하였다. 예를 들어 인위적 긴장감을 이용하여 극적인 집중효과를 유도할 수 있었으나, 이를 과감히 지양하여 작품에 자연스러움과 여유로움이 묻어날 수 있게 하였다. 또한 일차적으로 변형시킨 도자기 모양의 형상을 통해 한국인의 온화한 정서와 곡선의 아름다움을 다채로운 색채이미지로 해석해 보았다. 심상이란 일차원적 소재에 있어서만 인식되는 것이 아니다. 다양한 색의 다채로운 표현으로 인간본연의 환상과 환희, 다시 말해 원초적 감성에 자석처럼 이끌리는 자아를 발견하는 것이야말로 어쩌면 인간이 필연적으로 인식하여야 할 당위성을 확보하게 되는 것이다. 이를 인식하고, spot과 spot을 연결하는 구조를 통해 대상간의 접근성을 유기적으로 통합하여 구체화시키는 것이야 말로 필자가 추구하는 미적가치와 상응함은 사세고연한 일이다. 이러한 접근으로 심상 속의 회화적 표현연구는 복잡한 현재 우리들의 삶 속에서 자신의 발견과 표현에 대해 진지한 고민을 하고 새로움을 안고 출발하는 아름다운 여정이 될 것이라 확신한다.