Effect of an Endoplasmic Reticulum Retention Signal Tagged to Human Anti-Rabies mAb SO57 on Its Expression in Arabidopsis and Plant Growth

Song, Ilchan,Lee, Young Koung,Kim, Jin Wook,Lee, Seung-Won,Park, Se Ra,Lee, Hae Kyung,Oh, Soyeon,Ko, Kinarm,Kim, Mi Kyung,Park, Soon Ju,Kim, Dae Heon,Kim, Moon-Soo,Kim, Do Sun,Ko, Kisung Korean Society for Molecular and Cellular Biology 2021 Molecules and cells Vol.44 No.10

Transgenic Arabidopsis thaliana expressing an anti-rabies monoclonal antibody (mAb), SO57, was obtained using Agrobacterium-mediated floral dip transformation. The endoplasmic reticulum (ER) retention signal Lys-Asp-Glu-Leu (KDEL) was tagged to the C-terminus of the anti-rabies mAb heavy chain to localize the mAb to the ER and enhance its accumulation. When the inaccurately folded proteins accumulated in the ER exceed its storage capacity, it results in stress that can affect plant development and growth. We generated T₁ transformants and obtained homozygous T₃ seeds from transgenic Arabidopsis to investigate the effect of KDEL on plant growth. The germination rate did not significantly differ between plants expressing mAb SO57 without KDEL (SO plant) and mAb SO57 with KDEL (SOK plant). The primary roots of SOK agar media grown plants were slightly shorter than those of SO plants. Transcriptomic analysis showed that expression of all 11 ER stress-related genes were not significantly changed in SOK plants relative to SO plants. SOK plants showed approximately three-fold higher mAb expression levels than those of SO plants. Consequently, the purified mAb amount per unit of SOK plant biomass was approximately three times higher than that of SO plants. A neutralization assay revealed that both plants exhibited efficient rapid fluorescent focus inhibition test values against the rabies virus relative to commercially available human rabies immunoglobulins. KDEL did not upregulate ER stress-related genes; therefore, the enhanced production of the mAb did not affect plant growth. Thus, KDEL fusion is recommended for enhancing mAb production in plant systems.

Effect of Peanut Seed Orientation on Germination, Seedling Biomass, and Morphology in an Oak Tree Sawdust Cultivation System

Kisung Ko,Junsik Ahn,Ilchan Song,Dongjae Kim,Joon Chul Lee,Sungkwon Moon,Sooncheol Myoung 한국원예학회 2017 한국원예학회 학술발표요지 Vol.2017 No.10

주유소 내 부대시설 화재발생시 복사열에 따른 주유설비 안전거리에 관한 연구

김기성(Kisung Kim),이상원(Sangwon Lee),송동우(Dongwoo Song) 한국가스학회 2021 한국가스학회지 Vol.25 No.6

최근 주유취급소 내의 건축물에 편의점, 자동차 수리점 등의 부대시설이 증가하고 있다. 이에 따라 부대시설에서 화재 발생 시 고정주유기에 미치는 화재의 영향을 알아보고 현행 규정의 타당성을 검증할 필요성이 있다. 본 논문에서는 부대시설에서의 화재 시 고정주유기까지의 이격거리에 따라 미치는 복사열의 영향을 알아보는 연구를 진행하였다. FDS 5.5.3을 사용하여 화재모델링을 하였고, 화원의 크기는 실규모 화재평가 장치를 이용한 사무실의 연소실험을 통한 단위면적당 열방출율을 입력하였다. 화원과 고정주유기의 이격 거리는 ‘위험물안전관리법 시행규칙 별표13 주유취급소의 위치⋅구조 및 설비의 기준’에서 명시하는 2 m 부터 이격거리를 1 m 간격으로 늘여가며 10 m까지 화원의 크기를 Case A의 경우 512.41 kW/m², Case B의 경우 250 kW/m² 로 설정하여 총 13개의 시나리오를 구성하여 연구를 진행하였다. 시뮬레이션을 통해 2 m의 이격거리는 부대시설의 화재 시에 주유기의 안전성을 확보하지 못하는 것으로 나타났으며 최소 4m 이상의 이격거리에서 복사열이 피해를 미치지 않는 것으로 나타났다. 이에 따라 부대시설의 화재 발생시 복사열을 차단할 수 있는 설비가 필요하며 부대시설의 용도를 제한하거나 건축물과 고정주유기의 이격거리를 다시 제고하는 방안이 필요할 것이다. Various research has been done on fires and explosions at gas stations at home and abroad. However, only studies of off-site damage in the event of fire at the gas station were conducted, and research on fire at the auxiliary facilities in the gas station was insufficient. The gas station is a place where anyone can easily access dangerous goods. As the risk of fire increases due to the recent increase of auxiliary facilities such as convenience stores and car repair shops in gas stations, it is important to detect the effects of fire on the main oil refinery in case of fire and to verify the validity of existing regulations. In this thesis, we conducted a study to find out the effect of radiation heat on the separation between fixed and fixed oil reactors in the event of fire at an auxiliary facility. Simulation was modelled using FDS 5.5.3 Version, and the size of the fire source was configured with 13 fire assessment devices and the heat emission rate per unit area was entered. Simulation shows that the separation distance of 2 m does not secure the safety of the gas pump in the event of fire at the auxiliary facilities, and radiation heat does not damage at the separation distance of at least 4 m. Accordingly, facilities that can block radiant heat in the event of fire at auxiliary facilities, and measures to limit the use of auxiliary facilities or to re-impose the separation between buildings and fixtures will be needed.

Expression of Anti Colorectal Cancer Immunotherapeutic Monoclonal Antibody in Transgenic Plant

Mira Song,Mi-Hyun Ahn,Eun-Yi Oh,Zhe Lu,Da-Young Park,Jun-Hyung Kim,Misun Park,Arshad Jamal,Kisung Ko 한국원예학회 2008 한국원예학회 기타간행물 Vol.- No.-

Effects of ER retention motif on expression, glycosylation, and function of anti-cancer and anti-virus monoclonal antibodies in transgenic Arabidopsis

Ilchan Song,Young Koung Lee,kisung Ko 한국당과학회 2021 한국당과학회 학술대회 Vol.2021 No.07

Expression, Glycosylation, and Function of an Anti-rabies Virus Monoclonal Antibody in Tobacco and Arabidopsis Plants

Ilchan Song,Sol-Ah Park,Dalmuri Han,Hae Kyung Lee,Hyun Joo An,Kisung Ko 한국원예학회 2017 한국원예학회 학술발표요지 Vol.2017 No.10

새만금 농생명용지 지하배수암거의 효과분석

송재도 ( Jaedo Song ),손재권 ( Jaegwon Son ),최진규 ( Jinkyu Choi ),고남영 ( Namyoung Goh ),이기성 ( Kisung Lee ),이석주 ( Sukju Lee ),고대희 ( Daehee Goh ) 한국농공학회 2017 한국농공학회 학술대회초록집 Vol.2017 No.-

새만금 종합개발계획은 농업용지, 산업용지 등 8개 용지체계로 구분하였으며, 최근 농생명용지, 산업연구용지 ,국제교류용지 등 6개 용지체계로 변경하였다.(새만금 개발청, 2014) 농생명용지는 당초 수출농업전진기지, 친환경 고품질 첨단농산업기반, 녹색성장 농업생태관광 기반구축, R&D 및 농업서비스 지원 등 4대기능으로 설정하였고, 세부 13개 용지로 구분하여 첨단수출농업의 전진기지 육성, 농산업의 트랜드 변화에 대응 등을 구상하고 있다. 이러한 여건 속에서 새로이 조성되는 새만금 농생명용지에 대한 활용에 있어 관심이 집중되고 있는 상황이며, 토지이용계획에 있어 전작, 원예, 범용화 단지 조성을 계획하고 있다. 그러나, 간척지는 특성상 장기간 해수에 잠겨 있고, 경사가 완만하며 토양특성이 미사질 양토 계열로 원활한 배수가 이루어지지 않아 제염 및 작물생육에 어려움이 있어 작물재배를 위한 농지를 효율적으로 정비할 수 있는 하드웨어적 측면의 대책이 필요하다. 이에 본 연구에서는 새만금 농생명용지 5공구 내 지하배수암거의 시험시공에 따른 지하침출수량, 침출수의 염분농도 변화 등을 모니터링하여 지하침출 및 배수효과를 분석하였다.

연소 해석법을 이용한 Idle Irregular Shake 저감 연구

송재은(Jaeeun Song),이기성(Kisung Lee),김영권(Youngkwon Kim),윤재원(Jaewon Youn) 한국자동차공학회 2010 한국자동차공학회 부문종합 학술대회 Vol.2010 No.5

This study summarizes findings from recent efforts to better understand idle quality, including lumpiness (erratic shake). It combines new observations with previously known factors to provide a more complete view of the idle shake mechanism and the perception of the shake by occupants. The following discussion demonstrates the system nature of idle quality. Much of the data were processed by the combustion analysis method, which splits the idle vibration into its periodic and random components. This partitioning, coupled with several key observations of experimental data by various personnel, has led to a better understanding of idle lumpiness. Combining the combustion analysis method with cylinder pressure data also led to the calculation of the combustion torque spectra (random and periodic components) and to in-situ Frequency Response Functions. These measures quantify the engine’s combustion excitation, and the related vehicle sensitivities, respectively. Their characteristics explain trends first seen in the combustion analysis results. Using these techniques, an allocation method is described to ensure attainment of good idle quality.

Effect of Peanut Seed Orientation on Germination, Seedling Biomass, and Morphology in an Oak Tree Sawdust Cultivation System

Ahn, Junsik,Song, Ilchan,Kim, Dongjae,Lee, Joon Chul,Moon, Sungkwon,Myoung, Sooncheol,Ko, Kisung Korean Society of Horticultural Science 2017 원예과학기술지 Vol.35 No.4

We performed seed germination tests to investigate the effects of seed sowing orientation on germination viability on peanut (Arachis hypogaea L.) sprouts. Specifically, we assessed the influence of seed sowing orientation on germination rate, seedling weight, and seedling length, as well as the seedling vigor index. The seeds were sown in oak tree sawdust at 3.0 cm depth. Four seed orientations were tested: vertical with the hypocotyl end down, vertical with the hypocotyl end up, horizontal with the hypocotyl end down, and horizontal with the hypocotyl end up. The mean seed germination percentages of the four seed orientations were significantly different (p < 0.01) and ranged from 25 to 91.7%. The vertical orientation with hypocotyl-end-down and hypocotyl-end-up orientations showed the highest (91.7%) and lowest (25%) germination rates, respectively. The vertical orientation with the hypocotyl end down produced the heaviest (4.9 g) seedlings and the longest hypocotyls (4.65 cm). This orientation also produced the longest true leaf + epycotyl (2.15 cm) and had the highest seedling vigor index (197.1). The seedlings had a straight growth pattern, whereas seedlings from seeds sown with the hypocotyl up had an awkward plumular hook shape. Taken together, to produce peanut sprouts, we recommend placing the seeds vertically with the hypocotyl end down because this orientation leads to a high germination rate, high biomass production, and high overall seedling quality.

형질전환식물체 세대 간 항암단일항체 발현안정성

송미라(Mira Song),고기성(Kisung Ko) 한국원예학회 2007 원예과학기술지 Vol.25 No.3

본 실험은 형질전환된 담배로부터 세대가 거듭되더라도 항암 항체 유전자의 전달과 단백질의 발현에 변화가 없음을 확인하고자 수행되었다. 항암 항체 mAb CO17-1A를 발현하는 형질전환담배로부터 지속적인 자가수정을 통하여 얻은 씨앗에서 발아한 3세대와 4세대의 식물을 이용하였다. 형질전환된 담배 씨앗(3세대와 4세대)이 kanamycin이 포함된 배지에서 100% 발아 및 생장하는 것을 확인하였다. 이로써 3세대와 4세대 식물이 kanamycin에 내성이 있음을 알 수 있었으며, homozygous line임을 확인할 수 있었다. 약 16주 정도 키운 후 3세대와 4세대의 잎으로부터 genomic DNA를 추출하여 PCR을 수행한 결과 3세대 및 4세대의 식물에서 mAb CO17-1A의 중쇄(1.4kb)와 경쇄(0.7kb) 유전자가 모두 증폭됨을 관찰하였다. 즉, 세대가 거듭되더라고 유전자가 변이되지 않으며 안정하게 전이됨을 확인할 수 있었다. 또한 세대 번식을 통한 항체 단백질의 발현량을 확인하고자 3세대 및 4세대 식물의 잎으로부터 단백질을 추출하여 immunoblot을 수행한 결과 3세대 및 4세대 식물에서 중쇄(50kDa) 단백질이 동일한 density를 보였다. 이를 통해 세대가 거듭되더라도 항체를 생산하는 발현량에는 변화가 없음을 알 수 있었다. 위의 모든 결과를 바탕으로 형질전환 된 식물의 self-fertilization을 통하여 homozygous line의 세대번식이 이루어지며, 세대번식이 지속되더라도 그 유전자 및 발현량에는 변화가 없음을 확인할 수 있었다. Recent plant biotechnology and advanced molecular immunology have established plant biopharming system as an alternative way to produce immunotherapeutic monoclonal antibodies. This plant production system is considered to have several advantages in safety and economical issues compared to other production systems. In this study, we confirmed stable gene insertion and expression of anti-cancer monoclonal antibody (mAb CO17-1A) in transgenic plants through subsequent generation. T₃ and T₄ generation of transgenic seeds were obtained by self-fertilization of homozygous transgenic line of T₂ and T₃ generation, respectively. T₃ and T₄ seedlings showed 100% of germination and survival rate in MS media containing kanamycin whereas non-transgenic seedlings did not survive. PCR analysis showed that stable gene insertion of heavy and light chains of mAb CO17-1A in T₃ and T₄ transgenic plants. Immunoblot with densitometric analysis confirmed stable expression of mAb CO17-1A in both T₃ and T₄ transgenic plants. These results suggested that transgenes and expression of anti-cancer mAb CO17-1A are stably transferred and expressed in transgenic plants through subsequent generation by self fertilization, which is applicable to generate seed bank of homozygous transgenic line.