Comparison of Total Soluble Protein in Various Horticultural Crops and Evaluation of Its Quantification Methods

송일찬,김도선,김미경,Arshad Jamal,황경아,고기성 한국원예학회 2015 Horticulture, Environment, and Biotechnology Vol.56 No.1

Various horticultural crops have been considered as potential heterologous expression systems for plant-made recombinant pharmaceutical proteins. However, there is little information concerning the total soluble protein (TSP) levels of major horticultural crops. Ten major horticultural crops-Chinese cabbage, broccoli, garlic, onion, cabbage, bunching onion, cucumber, zucchini, radish, and carrot, along with tobacco and Arabidopsis, were selected to investigate the TSP levels in their individual tissues. In tobacco, SDS-PAGE assay showed that leaf tissues had stronger protein bands than stem tissues, and freshly harvested samples had slightly stronger band density than the -70℃ frozen samples, suggesting that fresh leaf should be used to measure the total soluble proteins without any protein loss or degradation. Bicinchoninic acid (BCA) protein assay revealed that among various horticultural crops, garlic (41.4 mg․g-1 FW), broccoli (21.9 mg․g-1FW), and Chinese cabbage (11.9 mg․g-1 FW) had the highest TSP levels suggesting that these horticultural crops could be good candidates for plant molecular biofarming to produce highly valuable recombinant proteins. The inner clove tissue in garlic, the flower tissue in broccoli, and the green leaf tissue in Chinese cabbage showed the strongest protein band density as compared to other tissues. The TSP of Arabidopsis tissues was quantified by SDS-PAGE, BCA, and Nano-drop methods. In general, the middle leaf tissue showed the highest TSP levels. To evaluate TSP levels of various horticultural crops, these three different methods were compared. The correlation and regression analyses between SDS-PAGE and BCA, and SDS-PAGE, and Nano-drop suggested that there were significant correlations between SDS-PAGE and BCA protein assays as compared to SDS-PAGE and Nano-drop assays, indicating that BCA assay is reliable to quantify TSP levels. In conclusion, the TSP levels varied depending on the horticultural crops and their tissue types, and BCA assay could be applied to quantify the TSP.

차량번호판 경매제도에 대한 상해시 시민사회의 반응

송일찬 대한교통학회 2019 대한교통학회 학술대회지 Vol.80 No.-

1978년 덩샤오핑이 개혁개방을 선언하기 직전, 중국의 도시화 비 율은 17.9%에 불과하였다. 이후 중국은 빠른 경제성장을 이루었고 현재는 14억 인구 중 도시에 살고 있는 인구가 절반이다. 그 결과 인구, 주택, 환경과 같은 중국의 온갖 사회문제가 도시라는 공간을 통해 압축적으로 드러나고 있다. 이 중 가장 심각한 문제는 교통체 증이다. 상해는 이러한 교통체증문제를 해결하기 위해 매달 경매를 통해 번호판을 부여하는 정책을 1994년부터 실시하고 있다. 이 정책은 효과적인 것처럼 보인다. 예를 들어 2004년에는 베이징과 상 해는 각각 2백만 대의 차량이 등록되어 있지만 현재 베이징은 600 만대인데 반해 상해는 360만대이다. 이처럼 경매제를 통해 차량수를 제한하고 교통 인프라에 연평균 100억 위안 이상을 투자하고 있지 만 상해의 교통체증 문제는 여전히 해결되지 않고 있다. 또한 구매 수요에 비해 훨씬 적은 차량쿼터를 경매를 통해 배분하다보니 경매 에서 입찰된 번호판 가격은 차 한대 값보다 더 비싸다. 이로 인해 차량 구매 자체가 경제력에 의해 제한되고 형평성 문제까지 제기된다. 교통정책을 보면 도시를 알 수 있고, 도시를 알면 그 사회를 알 수 있다. 도시의 인프라 설비가 늘어난다 해도 차량소유 증가로 인 한 도로 수요를 따라가지 못하는 상해의 교통문제 상황과 해결과정 은 우리나라에도 시사점이 많다. 하지만 이러한 중국 상해 번호판 경매제도에 대한 연구는 물론이거니와 중국의 교통정책에 관한 연구 가 국내에서 이루어진 바가 거의 없다. 이 연구는 상해시민 217명을 대상으로 상해 차량 번호판 경매제도에 대한 정책 수용성을 컨조인트 방식으로 조사한 것이다. 이를 통해 상해 시민들이 정책 효용성측면에서는 번호판 경매제도를 긍정 적으로 인식하고 있지만 형평성과 같은 다른 측면에서는 부정적으로 인식하고 있는 정책 수용성의 양면적 성격을 살펴볼 수 있다. 이러 한 분석과정을 분산분석과 효용성을 비롯한 컨조인트 설문조사 분석 을 통해 하였다. 그리고 이러한 분석에 앞서 상해의 정치, 사회, 경 제적 맥락을 경매제의 역사적 맥락과 함께 살펴보고, 이 맥락 속에 서 드러나는 문제점을 컨조인트 설문조사 결과와 함께 분석하여 정 책적 시사점을 제시하고자 한다. 이 정책의 대중적 수용성을 개선하기 위해서는 다음과 같은 세 가지 권장 사항을 제시한다. 현재 시행하고 있는 정책을 경매제에 대한 세분화를 통해 차량 쿼터제를 분류할 필요가 있다. 뿐만 아니 라 외부차량에 대한 단속이나 인구에 대한 실질적인 억제 정책 등 관련 정책에 대한 집행력을 강화시킬 필요가 있다. 마지막으로 차량 수 공급제한뿐만 아니라 차량사용 제한을 위한 혼잡통행료 부과 등을 통해 도심 내 차량 사용을 제한하는 정책을 강화할 필요가 있다.

Expression and in vitro function of anti-cancer mAbs in transgenic Arabidopsis thaliana

송일찬,강양주,김대헌,김미경,고기성 생화학분자생물학회 2020 BMB Reports Vol.53 No.4

The anti-colorectal cancer monoclonal antibody CO17-1A (mAb CO), which recognizes the tumor-associated antigen EpCAM, was expressed in transgenic Arabidopsis plants. PCR and western blot analyses showed the insertion and expression of heavy chain (HC)/HC fused to the KDEL ER retention modif (HCK) and light chain (LC) of mAb CO and mAb CO with HCK (mAb COK) in Arabidopsis transformants. Both plantderived mAbP CO and mAbP COK were purified from a biomass of approximately 1,000 seedlings grown in a greenhouse. In sandwich ELISA, both mAbP CO showed a slightly higher binding affinity for the target, EpCAM, compared to mAbM CO. In cell ELISA, both mAbsP COs showed binding affinity to the human colorectal cancer cell line SW480. Furthermore, mAbM CO, mAbP CO, and mAbP COK exhibited dose and timedependent regression effects on SW480 cells in vitro. In summation, both mAbP CO and mAbP COK, expressed in Arabidopsis, recognized the target antigen EpCAM and showed anti-proliferative activity against human colorectal cancer cells.

Expression, glycosylation, and function of an anti‑rabies virus monoclonal antibody in tobacco and Arabidopsis plants

송일찬,Sol‑Ah Park,Dalmuri Han,Hae Kyung Lee,안현주,고기성 한국원예학회 2018 Horticulture, Environment, and Biotechnology Vol.59 No.2

Plants have emerged as one of the most attractive systems for producing human therapeutic proteins against viral diseases. These include diagnostic reagents, vaccines, and antibodies. This process is known as molecular biofarming. The objective of this study was to develop and evaluate tobacco and Arabidopsis as plant platforms for producing human anti-rabies monoclonal antibody (mAb). Both tobacco and Arabidopsis transgenic plants were generated by Agrobacterium-mediated transformation. Purification of mAb SO57K from each plant was performed with ammonium sulfate-mediated precipitation and protein A affinity columns. SDS–PAGE analysis showed that the purity of mAb SO57K obtained from each transgenic plant was similar, whereas Arabidopsis showed approximately twofold greater protein expression than tobacco. The N-glycosylation was not significantly different between proteins from the two plant species, with both showing oligo-mannose glycan structures. The mAbs SO57 derived from both the model plants had similar neutralizing efficacy against target virus strain CVS-11. Taken together, tobacco and Arabidopsis are both promising platforms for producing a human anti-rabies mAb.

Effect of plasma-activated water on peanut seed germination and vegetative growth in a hydroponic system

송일찬,전형원,PRIATAMARYZAADITYA,Gayathri Subramani,고기성,Lee Young Koung 한국식물생명공학회 2023 Plant biotechnology reports Vol.17 No.4

Peanut sprouts have recently been reported to be rich in some nutrients, including vitamin C, resveratrol, antioxidants, and folate, and are consumed as functional food. However, the effect of indirect plasma treatment, such as plasma-activated water (PAW), on the germination rate and growth of peanut remains largely unknown. Here, we evaluated the effect of PAW produced using a surface dielectric barrier discharge (SDBD) device on germination and vegetative growth in peanut. SDBD-generated PAW had significantly higher nitrate content than tap water (TW). Compared with the TW group, PAW treatment increased the germination rate of peanut seeds by more than twofolds at 3 days after planting; however, there was no significant difference in final germination rate between the PAW and TW groups at 7 days after seeding. In addition, PAW treatment significantly improved biomass-related morphological traits in peanut sprout compared with TW treatment. At the cellular level, PAW treatment increased cell size in the hypocotyl in both the horizontal (115%) and longitudinal (209%) directions, resulting in increased hypocotyl growth. Moreover, histological analysis showed that PAW treatment significantly enlarged parenchyma cell size in hypocotyl pith tissue but did not affect cell number. Overall, these results showed that PAW treatment improved seed germination and growth by enlarging parenchyma and epidermal cell size in hypocotyl, resulting in increased biomass in peanut sprouts. QRT–PCR results showed that auxin response and growth-related genes (AhARF5, AhIAA13, SAUR32, AhGI-13, AhGI-75, AhCUC3-12, and AhCUC3-65) were upregulated in the PAW treated condition at 6 days compared to the TW treated sprouts.

중국 대도시의 고속철도역 개발에 관한 정책 네트워크 간 갈등 : 광저우남역(广州南站)을 중심으로

송일찬 대한교통학회 2021 대한교통학회 학술대회지 Vol.85 No.-

중국은 지난 10년 동안 고속철도시스템에 1조 달러 이상을 투자하여 빠른 속도로 고속철도망을 확장했다. 그 결과 중국은 일본, 독일, 프랑스 등 기존의 고속철도 국가들을 압도하는 전세계 2/3가 넘는 3만5000km를 보유하고 있다. 시간당 200km이상의 속도로 운행되는 고속철도망은 중국 전역의 주요도시를 연결하여 중국사회의 이동개념과 생활을 완전히 바꾸었고 중국의 국가 주도 개발 모델의 핵심으로 상징되고 있다. 또한 대규모 고속철도 건설은 중국이 급속한 도시화가 이루어지고 있는 상황에서 도시공간 확대라는 새로운 방향을 제시하였다. 고속철도 역세권은 기존 도시개발과 다른 새로운 도시역세권 형성을 통한 발전이 이루어지게 되는 것이다. 하지만 이러한 발전의 이면에는 고속철도역 부지 선택과 도시 개발에 있어 중앙정부, 지방정부, 중국국영철도그룹(中國鐵路) 간의 이해득실에 따른 갈등, 예측 수요에 대한 부정확한 예측으로 인한 무질서한 개발 및 자원 낭비, 설비 보수 및 유지로 인한 부채부담 증가로 이어지고 있는 상황이다. 뿐만 아니라 고속철도 주변에 세워진 수많은 신도시들이 유령도시가 되가는 현상은 고속철도역 개발의 현실을 보여주고 있다. 이러한 문제점의 원인은 고속철도 효과에 대한 지방정부의 합리적인 이해부족에 비롯된 것이 사실이나, 보다 핵심적인 문제는 부지개발과 주변환경에 대한 이해관계자의 무질서한 참여로 인하여 각 주체가 별개의 정책 수립을 하고 있다는 점에 있다. 이 연구는 중국의 대표적인 대도시 중 하나인 광저우(广州)에 있는 고속철도 광저우남역(广州南站) 개발 및 건설 과정에서 발생한 문제점을 살펴보기 위하여 고속철도 관련해서 특수 이해관계자인 철도부문의 중국국영철도그룹(中國鐵路)를 추가하여 기존의 정부-시장-사회 간의 분석을 강화하고자 한다. 또한 철도 부지선정과 개발에 있어 중앙정부와 지방정부 간의 갈등을 통해 중국 도시개발정책 과정의 일면을 정책인류학적 관점에서 살펴보고자 한다.

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

안준식,송일찬,김동재,이준철,문성권,명순철,고기성 한국원예학회 2017 원예과학기술지 Vol.35 No.4

We performed seed germination tests to investigate the effects of seed sowing orientation ongermination viability on peanut (Arachis hypogaea L.) sprouts. Specifically, we assessed theinfluence 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 thehypocotyl end up, horizontal with the hypocotyl end down, and horizontal with the hypocotylend up. The mean seed germination percentages of the four seed orientations were significantlydifferent (p < 0.01) and ranged from 25 to 91.7%. The vertical orientation with hypocotylend-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 producedthe heaviest (4.9 g) seedlings and the longest hypocotyls (4.65 cm). This orientation alsoproduced 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 withthe hypocotyl up had an awkward plumular hook shape. Taken together, to produce peanutsprouts, we recommend placing the seeds vertically with the hypocotyl end down because thisorientation leads to a high germination rate, high biomass production, and high overall seedlingquality.

Low risk of pollen-mediated gene flow in transgenic plants under greenhouse conditions

김득수,송일찬,고기성 한국원예학회 2018 Horticulture, Environment, and Biotechnology Vol.59 No.5

There is concern about the potential for transgene contamination in wild type (WT) plants via pollen-mediated gene flow from transgenic plants. In this study, we investigate pollen-mediated gene flow using tobacco transgenic lines carrying recombinant protein GA733-FcK, which is a putative candidate for producing colorectal cancer vaccine. Transgenic and WT plants were grown in greenhouse, with WT plants placed 0.3, 1, 5, 10, and 20 m away from transgenic plants. Seeds were harvested from randomly selected WT plants and sown on germination media supplemented with or without kanamycin. At 30 days after sowing, none of the WT seedlings produced true leaves and roots when grown on selective media. Polymerase chain reaction analysis revealed that GA733-FcK and nptII genes were expressed in shoots of transgenic plants but not of WT plants. Expression of GA733-FcK and nptII proteins was also abolished in WT leaves when compared to that of transgenic plants. Our findings suggested that there is low risk of pollen-mediated gene flow from transgenic plants expressing GA733-FcK when grown in greenhouse conditions.

Effect of Tobacco Mosaic Virus (TMV) Infection on Expression and Purification of Therapeutic Vaccine Proteins in Transgenic Plants

Chunha Shin,송일찬,이예지,So-Hyeon Baek,김대헌,고기성 한국원예학회 2020 원예과학기술지 Vol.38 No.2

Plant molecular biofarming has been increasingly studied in recent decades. Many kinds ofrecombinant immunotherapeutic proteins have been produced in transgenic tobacco plants. However, tobacco mosaic virus (TMV) can damage tobacco plants and cause pathogenic symptoms,which affects plant biomass production. Cigarette sap solution has been used to infect TMV duringtobacco cultivation. In this study, we obtained TMV-infected transgenic plants expressing a prostatespecificantigen (PSA) fused to an IgG Fc with KDEL ER retention motif (PSA-IgG FcK). Thetypical TMV-associated symptoms appeared and increased on plant leaves 5 days after infection. mRNA expression levels and the presence of TMV were confirmed by RT-PCR and transmissionelectron microscopy analyses. The expression level and purity of the target protein were notsignificantly different between noninfected and infected transgenic plants. TMV was not found inthe purified protein samples from infected plants. Our study showed that TMV pathogen-infectedplant biomass can be harvested and processed to obtain therapeutic vaccine proteins.

Relationship between ganglioside expression and anti-cancer effects of a plant-derived antibody in breast cancer cells

추영국,주원석,송일찬,박세라,서상영,조진형,민성훈,김대헌,김지수,김순욱,박순주,고기성 한국식물생명공학회 2019 JOURNAL OF PLANT BIOTECHNOLOGY Vol.46 No.3

Production of therapeutic monoclonal antibodies (mAbs) using a plant platform has been considered an alternative to the mammalian cell-based production system. A plant-derived mAb CO17-1AK (mAbP COK) can specifically bind to various types of cancer cell lines. The target protein of mAbP COK is the epithelial cell adhesion molecule (EpCAM) highly expressed in human epithelial cancer cells, including breast and colorectal cancer cells. It has been hypothesized that its overexpression supports tumor growth and metastasis. A ganglioside is extended well beyond the surfaces of the various cell membranes and has roles in cell growth, inflammation, differentiation, and carcinogenesis. However, the regulation of EpCAM gene expression in breast cancers and the role of gangliosides in oncogenesis are unclear. Here, the purpose of this study was to determine the effects of mAbP COK on human breast cancer cell proliferation, apoptosis, and ganglioside expression patterns. Our results show that treatment with mAbP COK suppressed the growth of breast cancer cells and induced apoptotic cell death. It also upregulated the expression of metastasis-related gangliosides in breast cancer cells. Thus, treatment with mAbP COK may have chemo-preventive therapeutic effects against human breast cancer.