인간 상피성장인자 유전자 발현 유전자변형 콩의 생태계 침입성 평가

김도영(Do Young Kim),엄민식(Min Sik Eom),김혜진(Hye Jin Kim),백인순(In-Soon Pack),박정호(Jung-Ho Park),박기웅(Kee Woong Park),남경희(Kyong-Hee Nam),오성덕(Sung-Dug Oh),김재광(Jae Kwang Kim),서주석(Ju Seok Seo),김창기(Chang-Gi Kim) 한국잡초학회·한국잔디학회 2020 Weed & Turfgrass Science Vol.9 No.2

유전자변형(genetically modified, GM) 작물의 생태계 침입성 또는 잡초화 가능성은 환경 측면에서 주요 우려 사항의 하나이다. 인간 상피성장인자 유전자(egf, igf-1, trx)를 발현하는 GM 콩 3개 이벤트가 재배관리되지 않는 환경에서 지속할 수 있는지 여부를 평가하고, 토양에 매몰된 종자의 활력을 조사하기 위해 본 연구를 수행하였다. GM 콩과 비변형 콩종자를 2017년 6월에 파종하고 연구기간 동안 물과 병해충 및 잡초 관리를 하지 않은 채 시험구에 출현한 콩 식물체 수를 조사하였을 때 GM 콩과 비변형 콩 식물체는 생장하고 종자 생산도 하였지만 17개월까지만 개체군을 유지할 수 있었다. 콩 종자를 2017년 11월에 매몰한 뒤 4, 6, 8, 10개월 후에 활력을 조사하였다. GM 콩과 비변형 콩 종자 모두 월동은 가능했지만, 모든 종자는 6개월 이내에 활력을 잃었다. 이와 같이 GM 콩과 비변형 콩 모두 정착에 성공하지 못했으며, 수명이 짧은 콩 종자는 일시적으로만 토양종자은행을 형성하였다. 따라서 3개 GM 콩 이벤트가 생태계에서 침입식물 또는 잡초가 될 가능성은 거의 없을 것으로 판단된다. The invasiveness or weediness of genetically modified (GM) crops is considered as one of the major environmental concerns regarding the cultivation of these crops. This study was conducted to evaluate whether three events of GM soybeans expressing human epidermal growth factor gene (egf , igf-1 , or trx ) can persist in the environment without cultivation management and to determine the viability of their seeds buried in soil. Seeds of GM and non-GM soybean were sown in June 2017, and the number of soybean plants grown in the plots was counted. During the study period, water and pest management were not applied in the plots. Both GM and non-GM soybean plants were able to grow and produce seeds; however, they maintained their populations only for up to 17 months. The GM and non-GM soybean seeds were buried in November 2017, and their viability was measured 4, 6, 8, and 10 months after seed burial. Both GM and non-GM soybean seeds were able to overwinter. However, they lost viability within 6 months. GM and non-GM soybeans could not be successfully established in the environment, and short-lived soybean seeds could form only a transient soil seed bank. Therefore, GM soybean events are highly unlikely to become invasive plants or weeds in the ecosystem.

GM 콩 DNA와 단백질의 안정성에 대한 열, 압력 및 산 처리의 영향

백인순(In-Soon Pack),정순천(Soon-Chun Jeong),윤원기(Won-Kee Yoon),박상규(Sangkyu Park),육은수(Eun-Soo Youk),김환묵(Hwan-Mook Kim) 한국식품과학회 2004 한국식품과학회지 Vol.36 No.4

Roundup Ready Soybean의 정량 검출에서 SYBR Green I을 이용하는 Real-Time PCR 조건의 분석

백인순(In-Soon Pack),정순천(Soon-Chun Jeong),이규화(Kyu Hwa Lee),윤원기(Won Kee Yoon),박상규(Sangkyu Park),김창기(Chang-Gi Kim),김환묵(Hwan Mook Kim) 한국독성학회 2005 Toxicological Research Vol.21 No.3

Concerns on the safety of living modified (LM) crops have led to mandatory-labeling legislation of LM crops in many countries including Korea. An real-time PCR method for quantification of LM Roundup Ready soybean (RRS) with the double-stranded DNA intercalating dye, SYBR Green I, is described. Pairs of primers that specifically PCR-amplify the targeted 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) and le1 gene sequences were selected by means of analysis of melting-curve plot of PCR products. Then, the specificity and general performance of the selected primer pairs were further increased by selecting optimum primer concentration by means of serial analyses of combination of different concentrations of each primer in the selected primer pairs. Using 1, 2, and 5% RRS test samples and applying the real-time PCR with SYBR Green I, mean values deviated from true values by 5.5 to 25%. The precision of the real-time PCR with SYBR Green I was comparable with that of the real-time PCR with TaqMan chemistry, which is widely used for the quantification of LM crops today. The results suggest that the real-time PCR with SYBR Green I, which does not require the high-cost dye labelling step for the initial primer or probe design, could be an alternative method for the quantification of LM Crops.

Bt 양배추가 복숭아혹진딧물의 발육과 기주선택에 미치는 영향

남기정,김영중,문두범,남경희,백인순,박정호,정순천,한지학,김창기,Nam, Ki Jung,Kim, Young-Joong,Moon, Doo-Bum,Nam, Kyong-Hee,Pack, In Soon,Park, Jung-Ho,Jeong, Soon-Chun,Harn, Chee Hark,Kim, Chang-Gi 한국응용곤충학회 2014 한국응용곤충학회지 Vol.53 No.2

해충저항성 유전자변형 작물은 대상 해충에 대한 방제 효과를 증진시킬 수 있는 효과적인 수단이 될 수 있지만, 생태계 내의 다른 비표적 생물에 부정적인 영향을 줄 가능성 또한 있기 때문에 상업적인 재배에 앞서 이에 대한 충분한 연구가 이루어져야 한다. 본 연구에서는 배추좀나방내성 유전자변형 Bt (Cry1Ac1)양배추가 비표적 곤충인 복숭아혹진딧물에 미치는 영향을 알아보기 위한 기초적인 실험으로서 기주선택실험 및 성장실험을 실시하였다. 기주선택실험에서, 두 개체씩의 Bt 양배추와 모본 양배추에 각각 10마리의 유시 성충(alate virginoparae)을 접종하고, 사흘간 생식(reproduction) 기회를 제공한 후 생산한 약충의 마리수를 측정한 결과, Bt 양배추와 모본 양배추에 낳은 약충의 수가 각각 $21.9{\pm}1.8$와 $22.5{\pm}2.2$로 비슷하였다. 성장실험에서, 갓 태어난 무시 약충(apterous virginoparae)을 Bt 양배추 및 모본 양배추를 기주로 하여 키웠을 때, 성충이 될 때까지 걸린 시간은 Bt 양배추와 모본 양배추에서 각각 $5.8{\pm}0.2$, $5.9{\pm}0.1$ 일(day)로 통계적으로 유의한 차이가 없었고, 개체군 생장률(rm) 또한 $0.7{\pm}0.1$, $0.8{\pm}0.1$로 유사하였다. 이상의 결과들로 볼때 Bt 양배추가 복숭아혹진딧물에 미치는 영향이 없거나 미미함을 알 수 있으나, 다양한 관점에서의 보완 연구가 더 필요하다. Transgenic crops that produce insecticidal toxins have a great potential for controlling target pest insects, but there is a growing concern about unintended influences on non-target species. In the present study, the preferences and performance of non-target green peach aphid, Myzus persicae (Sulzer), on transgenic cabbages (Brassica oleracea) that produce Bt toxin (Cry1Ac1) and untransformed control plants were investigated as a part of risk assessment. In a free-choice situation, the number of nymphs larviposited by 10 winged adults over 3 days was $21.9{\pm}1.8$ and $22.5{\pm}2.2$ on transgenic and the control plants, respectively, indicating that the aphids did not discriminate between the two types of plants. In a performance assay, the development time (D) and intrinsic rate of increase (rm) of wingless aphids reared on transgenic and control plants were also similar (D, $5.8{\pm}0.2$ and $5.9{\pm}0.1$ (days) and rm, $0.7{\pm}0.1$ and $0.8{\pm}0.1$, for transgenic and control plants, respectively). These results suggest that M. persicae is not significantly affected by transgenic Bt cabbage.

AtCYP78A7 과발현 환경스트레스 내성 형질전환 벼의 단백질 진단 키트 개발

남경희(Kyong-Hee Nam),박정호(Jung-Ho Park),백인순(In-Soon Pack),김호방(Ho Bang Kim),김창기(Chang-Gi Kim) 한국생명과학회 2018 생명과학회지 Vol.28 No.7

본 연구는 시토크롬 P450 단백질을 암호화하는 애기장대 유래의 AtCYP78A7을 과발현하는 형질전환 식물체로 부터 AtCYP78A7 단백질을 특이적으로 인식하는 단일큰론 항체의 제조와 그 항체를 AtCYP78A7 단백질과 접촉시켜 항원-항체 복합체 형성을 검출함으로써 AtCYP78A7 단백질을 효소면역학적(ELISA) 방법으로 검출하는 진단키트를 개발하기 위하여 수행하였다. 재조합한 GST-AtCYP78A7 단백질을 항원으로 사용하여 단일클론 항체를 분비하는 융합세포주를 제조한 후 비오틴화 및 페어링 테스트를 통해 포획항체와 검출항체를 선정하였으며, GST-AtCYP78A7 정제 단백질을 기준으로 일품벼, 화영벼, AtCYP78A7 과발현 벼(10B-5, 18A-4)의 용해물을 검출항원으로 사용하여 product test를 진행하였다. 그 결과 AtCYP78A7 단백질에 특이적으로 결합하는 4개의 단클론 항체(mAb 6A7, mAb 4C2, mAb 11H6, mAb 7E8)를 생산하였고, 포획항체 mAb 4C2와 검출항체 mAb 7E8-biotin의 조합으로 ELISA 키트를 개발하였다. 개발된 ELISA 키트를 이용한 벼 시료의 분석 결과 AtCYP78A7 과발현 벼는 전체 단백질 대비 AtCYP78A7 단백질의 비율이 0.1% 이상인 양성으로, 일품벼와 화영벼는 0.1% 미만인 음성으로 나타나 키트를 이용한 AtCYP78A7 단백질의 검출이 가능하였으며, 따라서 본 키트는 향후 AtCYP78A7를 과발현하는 형질전환 작물을 대상으로 하는 환경 모니터링 또는 인체 위해성 평가에 유용하게 활용될 수 있을 것으로 사료된다. Quantitative determination of the protein expression levels is one of the most important parts in assessment of the safety of foods derived from genetically modified (GM) crops. Overexpression of AtCYP78A7, a gene encoding cytochrome P450 protein, has been reported to improve tolerance to abiotic stress, such as drought and salt stress, in transgenic rice (Oryza sativa L.). In the present study, an enzyme-linked immunosorbent assay (ELISA) kit for diagnosing AtCYP78A7 protein including AtCYP78A7-specific monoclonal antibody was developed. GST-AtCYP78A7 recombinant protein was induced and purified by affinity column. Four monoclonal antibodies (mAb 6A7, mAb 4C2, mAb 11H6, and mAb 7E8) against recombinant protein were also produced and biotinylated with avidin-HRP. After pairing test using GST-AtCYP78A7 protein and lysate of rice samples, mAb 4C2 and mAb 7E8 were selected as a capture antibody and a detecting antibody, respectively, for ELISA kit. Product test using rice samples indicated that percentages of detected protein in total protein were greater than 0.1% in AtCYP78A7-overexpressing transgenic rice (Line 10B-5 and 18A-4), whereas those in negative control non-transgenic rice (Ilpum and Hwayoung) were less than 0.1%. The ELISA kit developed in this study can be useful for the rapid detection and safety assessment of transgenic rice overexpressing AtCYP78A7.

GM 작물의 내건성 검정과 위해성 평가를 위한 가뭄모의시설

남경희 ( Kyong Hee Nam ),고은미 ( Eun Mi Ko ),김도영 ( Do Young Kim ),백인순 ( In Soon Pack ),박정호 ( Jung Ho Park ),김창기 ( Chang Gi Kim ) 한국국제농업개발학회 2014 韓國國際農業開發學會誌 Vol.26 No.3

We designed and constructed a drought-simulation facility for use in producing droughttolerant transgenic crops and assessing their associated risks. This facility consists of soil storage containers and rainout shelters. Each container is equipped with an efficient drainage system so that proper drought conditions can be maintained by controlling water flow to and from the soil. The polyvinyl roof and sidewalls close automatically during rain events to exclude rainfall from plots within the containers but re-open under dry conditions. Water is supplied via sub-surface drip irrigation. The soil water content is monitored at 30-min intervals by sensors installed at depths of 10, 20, 30, and 40 cm. For testing its performance, the field within the facility was divided into two systems, well-watered and water-deficit. Three cultivars of rice were grown. During the experiment, the soil water content was relatively lower in the water-deficit system. Regardless of cultivar, the mean values for plant height; panicle length; grain number, weight, and width; 1000-grain weight; and overall yield were significantly lower in that system when compared with those parameters in the well-watered system.

유전자변형 양배추로부터 비유전자변형 모본으로의 유전자 이동성

김영중(Young-Joong Kim),남경희(Kyong Hee Nam),백인순(In Soon Pack),박정호(Jung-Ho Park),정순천(Soon-Chun Jeong),한지학(Chee Hark Harn),김창기(Chang-Gi Kim) 충남대학교 농업과학연구소 2014 농업과학연구 Vol.41 No.3

Understanding the gene flow from genetically modified (GM) crops to conventional crops is important to prevent and mitigate seed contamination caused by pollen-mediated gene flow. We conducted a field test to investigate the gene flow from diamondback moth resistant GM cabbage (Brassica oleracea var. capitata) containing cry1Ac1 gene, to a non-GM control line AD126. GM and non-GM cabbage plants were cultivated in the field and pollinated using Bombus terrestris under the nets during the flowering periods. After seeds were collected from non-GM plants, hybrids between them and the GM cabbages were screened by multiplex PCR targeting cry1Ac1 gene. Out of 878 germinated seedlings, 168 hybrids were found and the average gene flow frequency was 19.7%. Because cabbage is mainly pollinated by insect pollinators, large-scale field tests are needed to study gene flow of GM cabbage.