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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.
서부경남 재래시장을 대상으로한 유전자 재조합 대두의 모니터링
심원보,남백상,최주미,정순천,정덕화,Shim Won-Bo,Nam Baek-Sang,Choe Ju-Mi,Jeong Soon-Chun,Chung Duck-Hwa 한국환경보건학회 2006 한국환경보건학회지 Vol.32 No.2
The objective of this study was monitoring of genetically modified soybean by PCR and ELISA. We collected 60 soybean samples from the open markets located in Western Gyeongnam (Sacheon, Hamyang, Hadong, Sanchung, Uiryung, Geochang, and Hapcheon). A total of 60 soybeans was examined and 14 genetical modified soybean (GMS) were detected by PCR. The GMS rate of selling soybean in Uiryung, Hadong, Sacheon, and Hapcheon was 50.0%, 37.5%, 33.3% and 25%, respectively. The 7 of 14 GMSs were positive by ELISA and most of positive samples were below 3% GMS but 1 (Uiryung 1) of the positive samples was over the 3% which is maximum permit limit in Korea. These results mean that merchants of open market did not express for selling soybean mixed with GMS, so consumers did not recognize GMO. Therefore, we thought that education of GMO for merchant of open market need to recognize about GMO maximum permit limit.
유전자 변형 작물 성분 검출용 PCR Kit의 성능 평가 연구
윤시온(Si On Yun),정순천(Soon-Chun Jeong),윤원기(Won Kee Yoon),박상규(Sangkyu Park),문제선(Jae Sun Moon),이정현(Joung Hyun Lee),김환묵(Hwan Mook Kim) 한국독성학회 2004 Toxicological Research Vol.20 No.2
유전자변형작물의 이점과 잠재적 위해에 관한 다른 사회적 인식은 각국에서 다른 반응을 유발시켜왔다. 한국을 포함한 많은 국가는 새로운 규제를 제정하기 위해 부심히고 있다. 한국은 최근에 3% 이상의 유전자변형작물 혼입을 포함하는 모든 식품에 표시제를 실시하였다. 유전자변형작물 혼입을 신속하고 간편하게 검출하는 방법의 하나는 PCR에 의한 도입 DNA의 증폭이다. 이 목적을 위한 많은 PCR kit가 시판되고 있어, 본 연구는 이들 상업화된 유전자변형작물 검출 kit의 성능을 시험하였다. 그 결과 이들 15개 kit 중 6개는 안정성, 재현성 및 검출 한계의 측면에서 제작사 스스로 제시한 요구조건도 충족하지 못하여 이들 kit의 개발 및 생산 단계에서 품질관리에 문제점이 있음을 시사하였다. 본 시험은 또한 duplex와 triplex 검출 kit가 단일 PCR 반응에서 명백한 검출을 보장할지라도, monoplex 검출 kit의 검출 능력이 가장 높다는 것을 시사하였다. 또한, kit들은 콩과 옥수수 사이에서 다른 검출 한계를 보였다. 본 연구의 결과는 GM 작물의 재배, 국가간 이동, GM 작물을 사용한 식품 생산 과정의 모니터링 뿐만 아니라 GM 작물과 관련한 정부의 법규를 준수하기 위한 GM 작물의 혼입의 건전한 과학적 추적체계의 개발에 유용할 것이라 사료된다. The different social reflections about the benefits and the<br/> potential risks of genetically modified (GM) crops have evolved with different reactions in different countries. Many countries<br/> including Korea are working toward setting down new guidelines. Korea requires companies to label all food that contains more than 3% GM ingredients. One of the rapid and convenient detection methods of GM ingredients is amplification of the introduced DNAs by polymerase chain reaction (PCR). Many PCR kits for this purpose are commercially available. The objective of this study was to evaluate performance of commercialized GM crop detection kits. The results showed that 6 out of 15 kits tested did not meet the requirements even purposed by the manufacturers themselves in terms of stability, reproducibility, and detection limits, suggesting a potential quality control problem in their design stage or<br/> production line. The evaluation also suggests that, although the duplex and triplex detection kits allowed unambiguous detection in a single PCR reaction, the monoplex detection kits were the most sensitive to the detection of GM ingredients. The detection limits also differ between soybean and corn. Results from this study will be useful in the development of sound qualitative tracking systems of GM ingredients for monitoring throughout the cultivation of GM crops, their transboundary movement, and food production using GM grains as well as for complying with government guidelines associated with GM crops.
유전자 변형 작물 성분 검출용 PCR Kit의 성능 평가 연구
윤시온(Si On Yun),정순천(Soon-Chun Jeong),윤원기(Won Kee Yoon),박상규(Sangkyu Park),문제선(Jae Sun Moon),이정현(Joung Hyun Lee),김환묵(Hwan Mook Kim) 한국독성학회 2004 Toxicological Research Vol.20 No.1
The different social reflections about the benefits and the potential risks of genetically modified (GM) crops have evolved with different reactions in different countries. Many countries<br/> including Korea are working toward setting down new guidelines. Korea requires companies to label all food that contains more than 3% GM ingredients. One of the rapid and convenient detection methods of GM ingredients is amplification of the introduced DNAs by polymerase chain reaction (PCR). Many PCR kits for this purpose are commercially available. The objective of this study was to evaluate performance of commercialized GM crop detection kits. The results showed that 6 out of 15 kits tested did not meet the requirements even purposed by the manufacturers themselves in terms of stability, reproducibility, and detection limits, suggesting a potential quality control problem in their design stage or production line. The evaluation also suggests that, although the duplex and triplex detection kits allowed unambiguous detection in a single PCR reaction, the monoplex detection kits were the most sensitive to the detection of GM ingredients. The detection limits also differ between soybean and corn. Results from this study will be useful in the development of sound qualitative tracking systems of GM ingredients for monitoring throughout the cultivation of GM crops, their transboundary movement, and food production using GM grains as well as for complying with government guidelines associated with GM crops.
유전자 변형 탄저병 저항성 고추(PepEST gene)가 비표적 곤충인 복숭아혹진딧물(Myzus persicae Sulzer)에 미치는 영향
박지은,이훈복,김창기,정순천,윤원기,박기웅,이범규,김환묵,Park, Ji-Eun,Yi, Hoon-Bok,Kim, Chang-Gi,Jeong, Soon-Chun,Yoon, Won-Kee,Park, Kee-Woong,Lee, Bum-Kyu,Kim, Hwan-Mook 한국응용곤충학회 2007 한국응용곤충학회지 Vol.46 No.3
탄저병에 저항성을 가진 특정 유전자 esterase gene (PepEST)을 삽입한 유전자 변형 작물 PepEST (line 68) 고추가 비표적 생물체인 복숭아혹진딧물의 생장에 미치는 영향을 알아보기 위하여 복숭아혹진딧물 성충의 발육 및 산자수 등을 온도 $25^{\circ}C$, 상대습도 50-70%, 광주기 L16 : D8의 실험실 조건에서 조사하고 생명표를 작성하였다. 총 3회의 실험에서 순생산률($R_0$), 증가율($r_m$), 평균 재생산 기간(T), 총 생산량, 수명, 세대기간 등을 각각 산출하였다. 그 결과 수명은 세 번의 실험에서 각각 31일, 27일, 25일로 나타나 복숭아혹진딧물의 일반적인 평균 수명인 25-29일을 기준으로 유전자변형 고추와 모본 고추 모두에서 유의한 차이를 나타내지 않았다. 최초 산자일 또한 세 번의 실험 모두 유전자변형 고추와 모본 고추간에 차이를 나타내지 않았다(P>0.05). 반면, 총 산자수의 값과 Jackknife로 계산된 순생식률($R_o$)의 값은 유전자변형 고추보다 모본 고추에서 높은 결과를 나타냈다(P<0.05). 결과적으로 본 조사로 인해 Esterase gene PepEST를 삽입한 유전자 변형 작물이 복숭아혹진딧물의 산자수에 영향을 미치는 결과를 가져왔고, 유사한 방법의 실험을 야외에서 수행할 필요가 있을 것으로 본 연구는 제안한다. To assess the environmental risks of transgenic chili pepper with PepEST gene on non-target organisms before it exposes to the agro-ecosystem environments, we conducted the three sets of green peach aphids (Myzus persicas S.) life table experiment under laboratory conditions (Temp. $25^{\circ}C$, R.H. 50-70%, Photoperiod L16 : D8) in series during 2005-2006. We measured the net reproductive rate ($R_0)^*$, the intrinsic rate of increase ($r_m$), the mean generation time ($T_c$), $fecundity^*$, life span, and reproduction period between non-transgenic chili peppers and transgenic chili peppers, respectively. The life span of green peach aphids from three sets was 31, 27, 25 days, and the period of life span was similar to the general average length of green peach aphids, 25-29 days. Although the first reproduction of transgenic pepper was similar to the non transgenic pepper (P>0.05), the fecundity and the net reproductive rate ($R_o$) by using Jackknife method of transgenic pepper were lower than those of non transgenic pepper (P<0.05). Conclusively, we observed the adverse effect from our results but we should execute further experiments to confirm the results at the fields with the similar way.
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.
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.