Effect of ER signal peptides (KDEL) on expression and function of monoclonal antibody in plant

Da-Young Park,Eun-Yi Oh,Zhe Lu,Yangkang So,Mi-Hyun Ahn,Mira Song,Kyung-Jin Lee,Doo-Byoung Oh,Youngkwan Kim,Kisung Ko 한국당과학회 2009 한국당과학회 학술대회 Vol.2009 No.1

Plant genetic engineering led to the production of plant-derived mAb (mAbP), which provides a safe and economically feasible alternative to the current animal expression systems. In this study, the expression level of mAbP SO57 between with and without Lys-Asp-Glu-Leu (KDEL) was compared in transgenic plant. PCR and PT-PCR analyse showed stable gene insertion transcription of heavy and light chain genes of mAbP SO57 with or without KDEL in plant, respectively. We validated expression of mAbP SO57 by western blot. Western blot showed the significantly higher expression level of mAbP SO57 with KDEL compared to without KDEL. Flow cytometry (FACS) analysis showed that the Fc domains of both purified mAbP and mammalian-derived mAb (mAbM) evidenced similar binding activity to the FcγRI receptor (CD64). High performance liquid chromatography (HPLC) analysed, glycosylation patterns of mAbP SO57 with or without KDEL. The mAbP SO57 with KDEL had glycan profile with both oligomannose type (47.6%) and golgi type (52.4%), while the mAbP SO57 without KDEL had only golgi type (100%) glycans. Neutralizing analysis with rabies virus CVS-11 showed the similar neutralizing activity between mAbP SO57 with and without KDEL. These results suggest the potential of mAbP SO57 for rabies immunotherapy, regardless of plant specific glycan structures.

林地利用에 따른 溪流水의 水質變化

具瀟瑛,鄭源玉,金弘洙,朴晋源,麻鎬燮 경상대학교 농과대학 부속연습림 2001 演習林硏究報告 Vol.- No.11

Table 2. Evaluation of stream water quality in three stands by Drinking water quality standard <원문참조> 임지이용에 따른 수질변화 및 오염정도를 구명하여 향후 비점오염원 관리를 위한 기초자료를 제공하고자 경남 진주시 정평리에 위치한 3개 임분(소나무, 밤나무Ⅰ, 반나무Ⅱ임분)에서 2000년 7월부터 9월까지 계류수의 수질변화를 분석하여 얻은 결과는 다음과 같다. 1. 세 임분의 계류수 pH는 소나무 임분(pH6.59)>밤나무 Ⅱ임분(ph 6.53)>밤나무 Ⅰ임분(pH 6.47)계류수 순으로 나타났다. 2. 전기전도도는 밤나무 Ⅰ임분>소나무 임분>밤나무 Ⅱ임분 계류수의 순으로 높게 나타났다. 3. 양이온은 세 임분의 계류수 모두 Ca^2+, Na^+, Mg^2+, K^+, NH_^+의 순으로 높게 나타났다. 4. 음이온은 소나무 임분과 밤나무 Ⅱ임분 계류수에서는 SO_4^2, Cl, NO_3의 순으로 높게 나타났고, 밤나무 Ⅰ임분 계류수에서는 NO_3, SO_4^2, Cl 의 순으로 높게 나타나 차이를 보였다. 5. 소나무, 밤나무 Ⅱ임분 계류수는 pH, NH_4+, NO_3, Cl, SO_4^2이 먹는 물 수질기준 범위내에 있었다. 그러나 밤나무 Ⅰ임분 계류수에서 5항목 중 NO_3는 먹는 물 수질기준을 넘어서는 범위에 있어 임지이용에 따른 하류수질의 오염이 우려되었다. 따라서 이러한 상류지역에서 오염원 차단을 위한 수질정화시설 등의 대책이 마련되어야 할 것으로 사료된다. This study was carried out to clarify the change characteristics of stream water quality by land-use of forest from July to September, 2000 in three stands(Pinus densiflora, Castanea crenata(Ⅰ), Castanea crenata(Ⅱ) stand) of Jeongpyeong-ri, Jinju-si, Gyeongnam. The mean pH of rainfall results in acid rain of 5.3. The pH of stream water in three stands was hugh in order of Pinus densiflora (pH 6.59), Castanea crenata(Ⅱ)(pH 6.53) and Castanea crenata(Ⅰ) stand(pH 6.47). The electrical conductivity of stream water was high in order of Castanea crenata(Ⅰ), Pinus densiflora and Castanea crenata(Ⅱ) stand. Cations of three stand in stream water high in order of Ca^2+, Na^+, Mg^2+, K^+ and NH_4^+. But anions of stream water in Pinus densiflora stand and Castanea crenata(Ⅱ) stand were high in order of SO_4^2, Cl and NO_3. In Castanea crenata(Ⅰ) stand cations of stream water were high in order of NO_3, SO_^2 and Cl. The level of pH, NH_4^+, NO_3, Cl and SO_4^2 of stream water in Pinus densiflora stand and Castanea crenata(Ⅱ) stand reached within the level of domestic use standard for drinking water But the level of NO_3 of stream water in Castanea crenata(Ⅰ) stand was higher than that of domestic use standard. Therefore, non-point sources like forest watersheds which are fertilizer application lands should be taken to the appropriate mitigation measures.

Effect of ER signal peptides (KDEL) on expression and function of anti-virus monoclonal antibody in plant

Da-Young Park,Yangkang So,Kyung-Jin Lee,Zhe Lu,Eun-Yi Oh,Kwang-Wok Min,Seul-Ki Lee,Jung-Hwan Lee,Mi-Hyun Ahn,Mira Song,Doo-Byoung Oh,Youngkwan Kim,Kisung Ko 한국당과학회 2009 한국당과학회 학술대회 Vol.2009 No.1

Plant genetic engineering has led to the production of plant-derived mAb (mAbP), which provides a safe and economically feasible alternative to the current antibody expression systems. In this study, the expression levels of mAbP SO57 with or without ER-signal peptides (Lys-Asp-Gly-Leu;KDEL) in transgenic tobacco plants were analysed in transgenic plant. PCR and Reverse Transcription-PCR analyses showed existence of heavy and light chain genes of mAb with or without KEDL and their transcription in plant, respectively. Western blot showed that the expression levels of mAbP SO57 with KDEL were significantly higher than that without KDEL. Flow cytometry analysis showed that the Fc domains of both purified mAbP and mammalian-derived mAb have similar binding activity to the FcγRI receptor. High-performance liquid chromatography analysis showed that the mAbP SO57 with KDEL had glycan profile with both oligomannose and golgi type, whereas the mAbP SO57 without KDEL had only golgi type glycans. Neutralizing analysis with rabies virus CVS-11 showed the similar neutralizing activity between mAbP SO57 with and without KDEL. These results suggest that the potential of mAbP SO57 for rabies immunotherapy is regardless of plant specific glycan structures.

SOD2와 NDPK2 유전자 집적에 의한 페튜니아의 아황산가스 저항성 증진

이수영(Su Young Lee),천경성(Kyeong-Seong Cheon),김소영(So Young Kim),권오현(O Hyeon Kwon),이혜진(Hye Jin Lee),김원희(Won Hee Kim),유봉식(Bong Sik Yoo) 한국원예학회 2016 원예과학기술지 Vol.34 No.1

유전자 집적에 의해 아황산가스 저항성 증진 페튜니아를 개발하고자 비생물적스트레스 저항성 유전자로 널리 알려진 SOD2 와 NDPK2유전자가 각각 도입된 SOD2 형질전환 계통 SOD2-2-1-1-35(T₄)[S(T₄)]와 NDPK2 형질전환 계통 NDPK2-7-1(T₂)[N7-1(T₂)]간 상호교잡을 실시하여 교잡 제1세대를 획득하였다. 교잡 제1세대 중 SOD2와 NDPK2유전자가 모두 집적된 개체 획득율은 32.1-73.0%이었고, SOD2와 NDPK2유전자가 모두 집적된 개체는 SOD2 또는 NDPK2유전자가 단독 도입된 개체에 비해 아황산가스 30ppm처리 피해율이 2.6-5.1배 낮아 아황산가스 저항성이 증진되었음을 확인하였다. 또한 아황산가스 저항성이 증진된 교잡 제1세대를 자가수분에 의해 세대진전 시켜 획득한 교잡 제2세대의 아황산가스 저항성 검정을 통하여 교잡 제1세대에서의 아황산가스 저항성이 후대에서도 안정적으로 발현함을 확인하였다. 또한, 광합성 효율 증진 조사를 통해 아황산가스 저항성이 증진된 SOD2와 NDPK2유전자 집적 후대가 아황산 가스에 대한 스트레스를 덜 받고 있음을 확인하였고, RT-qPCR분석을 통해 SOD2와 NDPK2유전자 집적 후대의 아황산가스 저항성 증진이 SOD2와 NDPK2 유전자 집적에 의한 효과임을 확인하였다. The aim of this study was to develop a transgenic petunia with enhanced resistance to sulfur dioxide (SO₂) gas by stacking two genes, SOD2 and NDPK2, which are both known to confer resistance to abiotic stresses. The first-generation hybrids (TF1) were obtained through reciprocal crosses between an SOD2-transgenic line SOD2-2-1-1-35(T₄)[S(T₄)] and an NDPK2- transgenic line NDPK2-7-1(T₂)[N7-1(T₂)]. Approximately 32.1-73.0% of the first-generation hybrids (TF₁) carried both SOD2 and NDPK2 genes. These hybrids showed 2.6 and 5.1 times less damage than hybrids carrying only SOD2 or NDPK2 genes, respectively, when they were treated with SO₂ gas at 30 ppm. This confirmed that the heterozygous hybrids were more resistant to SO₂ than the hybrids carrying either one of the resistance genes. Second-generation hybrids (TF₂) were obtained by selfing the TF₁ individuals. We confirmed the expression of the stacked genes in the TF₂ hybrids by phenotypic observation of their response to SO₂ gas at 30 ppm as well as using RT-qPCR and photosynthetic efficiency.

Interacelluar reprogramming of expression, glycosylation, and function of plant-derived anti-virus therapeutic monoclonal antibody

Da-Young Park,Kyung-Jin Lee,Yangkang So,Zhe Lu,Seulki Lee,Jung-Hwan Lee,Seung Han Oh,Yeon Soo Han,Doo-Byoung Oh,Youngkwan Kim,Kisung Ko 한국당과학회 2010 한국당과학회 학술대회 Vol.2010 No.1

Plant genetic engineering has led to production of plant-derived mAb (mAbP), which can provides a safe and economically effective alternative to the conventional antibody expression methods. In this study, the expression levels of mAbP SO57 with or without ER-retention peptide extensions signal (Lys-Asp-Glu-Leu; KDEL) in transgenic tobacco plants were analysed in transgenic plant. The expression levels of mAbP SO57 with KDEL were significantly higher than that without KDEL regardless of transcription level. mAbP SO57 with KDEL localized surround to the nucleus suggesting that the mAbP with KDEL is localized in ER. The mAbP without KDEL and mAbH had mainly Golgi type glycans, whereas the ER-localized mAbP with KDEL had glycan profile with both oligomannose type (47.6%) and Golgi type (52.4%). The Fc domains of both purified mAbP (with and without KDEL) and human-derived mAb (mAbH) had similar bingding activity to the Fcγ RI receptor (CD64). Both mAbP (with and without KDEL) had a shorter half-life than mAbH. However, both mAbP with and without KDEL was as effective at neutralizing activity of the rabies virus CVS-11 as the mAbH. These results suggest that ER localization of recombinant mAbP by KDEL reprograms glycosylation and enhances production of the functional antivirus therapeutic antibody in the plant.

HSS STI-CMP적용을 위한이중 패드의 최적화

서용진,박성우,김철복,정소용,이경진,김기욱,박창준 대불대학교 2002 大佛大學校大學院 硏究論文集 Vol.- No.1

As the device geometry shrinks to the deep submicron region, chemical mechanical polishing(CMP) planarization become a more essential technique of advanced ULSI process. Also, CMP process was required for the global planarization of inter-metal dielectric(IMD), inter-level dielectric(ILD) layers and interconnections with free-defect. Especially, the complete global planarization of IMD, ILD and interconnections can be achieved only with the CMP process. However, as the IMD and ILD layer gets thinner, several problems were found in the CMP process. It does have various problems such as dishing effect, torn oxide defects and nitride residues in oxide. So, it leads to severe circuit failure, which affects yield. In this paper, we studied the characteristics of polishing pad, which can apply STI-CMP process for global planarization of multilevel interconnection structure. Also, we investigated the effects of different sets of polishing pad, such as soft and hard pad. As an experimental result, hard pad showed center-fast type, and soft pad showed edge-fast type. Totally, the defect level has shown little difference, however, the counts of scratch was defected less than 2 on JR111 pad. Through the above result, we can select optimum polishing pad, so we can expect the improvement of throughput and device yield.

남자대학생의 흡연태도와 행동에 관한 연구

강경진,김정미,민혜영,안소진,오경숙,은경희,이현,이희선,홍주미,김수지 이화여자대학교 간호과학대학 2003 이화간호학회지 Vol.- No.37

There have been so many researches about problems of cigarettes and relation of smoking and heath since 1950's and generally known that smoking is one of the worst factor of increasing disease and mortality. No matter what such many proofs about various problems of smoking have been researched, smoking rate of current male collage students has not been decreased. Besides, most of the researches about cigarette use and its addiction have been focused on the adolescent or adult women smokers or adult male, not male collage student. Therefore, the subject of this study is about smoking attitude and smoking behavior of male collage students by general features. Smoking attitude was analyzed by modifying 'Guidelines for the content of tobacco smoking surveys of the general population'(WHO, 1982). Two hundred of male collage students participated to the distributed questionnaire for one week in May. The smoking attitude was categorized into duration of smoking and number of cigarette per day(CPD), and nicotine dependency, investigated by Fagerstrom Tolerance Questionnaire(FTQ) (Fagerstorm & Schneider, 1989). All statistics were performed by procedure of SAS according to number, percentage, mean, standard deviation, and ANOVA. The results are as follows: 1. Smoking attitude level was mean 25.73±5.15 marks, ranging from 13 to 39 marks of the total 40 marks. 2. Duration of cigarettes, number of cigarette per day, and nicotine dependency were mean 5.14±2.84 years, 12.88±6.85 pieces, and 4.78±1.69, respectively. 3. Smoking attitude showed significance according to general features such as marital state and allowance, but the significance according to marital state is not valuable because the numerical difference is so big between married one and single that they cannot be compared with each other. 4. Duration of smoking was significant according to general features such as age and allowance. 5. Smoking amount showed significance the factors of age only. 6. ll1ere was no significance in nicotine dependency according to general features. As a result, smoking attitude of male collage student in this study shows moderate level. Smoking attitude and duration is significant according to allowance, and smoking amount is significant according to age. Nicotine dependency according to general features is not significant but comparing with the preceding study about adult male, male collage students in this study are respectively more nicotine-dependent than adult male smokers. In conclusion, the right allowance usage strategies and effective heath education focused on the young are needed. In addition, helpful programs and information spreading strategies are necessary for establishing higher level of smoking attitude. At last, systemic regulations in daily living for ""anti-nico-dependent"" and perspective reforming of male university student are needed for reducing nicotine dependency.

서울 시내 4년제 남자 대학생의 우울과 흡연량, 흡연기간, 니코틴 의존도와의 관계

박지영,박소영,이미숙,백수진,신의경,예혜련,오승진,장윤정,조혜진,김수지 이화여자대학교 간호과학대학 2003 이화간호학회지 Vol.- No.37

Background: With reports on the relation between smoking and depression appearing since 1980, it has become clear that depression is one of the psychodynamics of smoking, though there has been little or no progress made in the study of whether or not there is indeed a relation between depression and nicotine dependence. Purpose: The purpose of the study was to investigate the effects of depression on the amount and period of smoking and nicotine dependence among university man student smokers. Method: In periodical examination between 14 and 21, May, 2003, using 170 university male. The CES-D(Center for Epidemiologic Studies Depression) Scale and the FTQ(Fagerstrom Tolerance Qustionnaire) were utilized to measure the level of depression and nicotine dependency. The amount of smoking was calculated based on the number of cigarette used. Result: Results indicated that depression had something to do with amount of smoking. Nicotine dependence also was related to depression but total value was relatively low. finally the period of smoking had nothing to do with depression. Conclusion: The results of this study revealed that the amount of smoking of university man student smokers was very high. Both the amount of smoking and nicotine dependence were affected by levels of depression. Accordingly psychiatric nursing access such as emotional support is needed to manage depression and decrease smoking.

간호실무에서 신봉이론(Espoused theories)에 대한 탐색적 연구

서문자,김혜숙,이은희,박영숙,조경숙,강현숙,임난영,김주현,이소우,조복희,이명하,지성애,하양숙,손영희,권성복,김희진,추진아 성인간호학회 2001 성인간호학회지 Vol.13 No.1

As a nursing practice involves nurses' actions in a specific context of health care, this study has focused on exploring the espoused theories in nursing practice within the action science perspectives, Espoused theories are the belief, principles, and rationale expressed by the practitioner as guiding her/his actions in a situation of practice. The data were analysed qualitatively and 25 elements of espoused theories of nursing action were identified and clustered into 6 categories. The 25 elements of espoused theories are as follows: The clinical nurse worked in wholistic and individual nursing, focussed on the patient's needed, comfort and supportive nursing (5 theories of nursing goal) ; excellent skills, knowledge based, assessment and data collection, explaining, educating or a scientific basis(6 theories of nursing intervention): advocacy, value oriented, treatment, account- ability and commitment(4 theories of nursing ethics) ; human respect. partnership, trust(3 theories of patient-nurse relationship) : knowledgable, accumulated clinical experiences and personally lived experiences. positive perspectives(4 theories of nurse), role of intervention. rewarding peer relationship(3 theories of situations) The above mentioned espoused theories are similar to that of nursing textbooks which students learned through basic nursing education and almost the same as the Acts ofa Nurse in Korean. However, we are doubtful whether nurses actually do as they think. Therefore. it is recommended to review the theories-in-use in order to and any discrepancies between the espoused theories and the reality of nursing actions

콘크리트 중력식댐의 내진성능 평가에 관한 연구

소진호,정영수 중앙대학교 건설환경연구소 2001 環境科學硏究 Vol.12 No.1

The objective of this study is firstly to frame up the seismic safety of concrete gravity dams. It is necessary to analyze seismic response and evaluate seismic performance off concrete gravity dams during earthquake. In this study, seismic damage and dynamic analysis of concrete gravity dams using SAP2000 program are performed. Additional dynamic water pressure due to earthquake was considered as additional mass for numerical seismic analysis. But, a further research will be necessary for the seismic stability of dams.