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형질전환 베타카로틴 강화 콩 계통 선발 및 도입유전자 특성 분석
친양(Yang Qin),권순종(Soon-Jong Kweon),정영수(Young-Soo Chung),하선화(Sun-Hwa Ha),신공식(Kong-Sik Shin),임명호(Myung-Ho Lim),권택(Taek-Ryoun Kwon),조현석(Hyun-Suk Cho),박순기(Soon Ki Park),우희종(Hee-Jong Woo) 한국육종학회 2015 한국육종학회지 Vol.47 No.2
The β-carotene biofortified transgenic soybean was developed recently through Agrobacterium-mediated transformation using the recombinant PAC (Phytoene synthase-2A-Carotene desaturase) gene in Korean soybean (Glycine max L. cv. Kwangan). GM crops prior to use as food or release into the environment required risk assessments to environment and human health in Korea. Generally, transgenic plants containing a copy of T-DNA were used for stable expression of desirable trait gene in risk assessments. Also, information about integration site of T-DNA can be used to test the hypothesis that the inserted DNA does not trigger production of unintended transgenic proteins, or disrupt plant genes, which may cause the transgenic crop to be harmful. As these reasons, we selected four transgenic soybean lines expressing carotenoid biosynthesis genes with a copy of T-DNA by using Southern blot analysis, and analyzed the integration sites of their T-DNA by using flanking sequence analysis. The results showed that, T-DNA of three transgenic soybean lines (7-1-1-1, 9-1-2, 10-10-1) was inserted within intergenic region of the soybean chromosome, while T-DNA of a transgenic soybean line (10-19-1) located exon region of chromosome 13. This data of integration site and flanking sequences is useful for the biosafety assessment and for the identification of the β-carotene biofortified transgenic soybean.
레스베라트롤 합성 GM 벼 검정을 위한 계통특이 마커 개발
우희종(Hee-Jong Woo),친양(Yang Qin),백소현(So-Hyeon Baek),여윤수(Yunsoo Yeo),신공식(Kong-Sik Shin),임명호(Myung-Ho Lim),조현석(Hyun-Suk Cho) 한국육종학회 2016 한국육종학회지 Vol.48 No.2
A variety of genetically modified (GM) crops have been developed in Korea. In these crops, the resveratrol-enriched transgenic rice plant (Agb0102) has moved ahead to generate the dossier for regulatory review process required for commercialization of GM crop. The resveratrol-enriched transgenic rice plant could be released to farmers for cultivation after national regulators have determined that it is safe for the environment and human health. Here, we developed a PCR-based DNA marker based on flanking sequences of transgene for the discrimination of resveratrol-enriched transgenic rice plant. This DNA markers will be useful for identifying of resveratrol-enriched transgenic rice plant, and can also be used to estimate transgene movement occurred by pollen transfer or seed distribution. Moreover, it is helpful for prompt screening of a homozygote-transgenic progeny in the breeding program.
연구보문 : 자연과학; 유전자변형 병저항성(OsCK1) 벼에 대한 PCR 검정
신공식 ( Kong Sik Shin† ),이진형 ( Jin Hyoung Lee ),임명호 ( Myung Ho Lim ),우희종 ( Hee Jong Woo ),친양 ( Yang Qin ),조현석 ( Hyun Suk Cho ) 한국국제농업개발학회 2013 韓國國際農業開發學會誌 Vol.25 No.3
Genetically modified (GM) crops generated by agricultural biotechnology are rapidly increasing and cultivated in many countries since they have delivered substantially agronomic, environmental, economic, and social benefits to farmers. The disease resistant transgenic rice, which expresses choline kinase gene (OsCK1), was developed. With the potential problems of safety, the detection method is required as an essential element for the GMO labeling system or GMO traceability of transgenic crops. In this study, gene, construct and event-specific primer pairs with 134, 306 and 243 bp amplicon, respectively, were used for PCR amplification of disease resistant (OsCK1) GM rice and no amplified product was observed from any other crops as templates. The limits of detection (LOD) of qualitative duplex PCR were 0.05% for OsCK1 GM rice using the event-specific primer pair CKRB32-1/02-2. For quantitative detection using TaqMan real-time PCR system, plasmid pSPSCKR as reference molecule was constructed, which contains rice endogenous gene sequence and 5`-junction DNA sequence of OsCK1 GM rice. The absolute detection limit of quantitative PCR method was around 10 copies for one plasmid molecule pSPSCKR. Thereafter, five mixed transgenic rice containing 0.5, 1, 3, 5 and 10% were quantified to evaluate the accuracy and precision of the established real-time PCR system. The bias and the relative deviations were all within the range of 20% for OsCK1 GM rice. These results demonstrate that the developed event-specific qualitative and quantitative PCR methods are acceptable for monitoring and traceability of GM rice.
우희종(Hee-Jong Woo),신공식(Kong-Sik Shin),임명호(Myung-Ho Lim),이진형(Jin-Hyoung Lee),친양(Yang Qin),박순기(Soon Ki Park) 한국육종학회 2015 한국육종학회지 Vol.47 No.1
The selectable marker-free rice plants containing mcry1Ac insecticidal gene isolated from Bacillus thuringiensis (Bt) were generated using a non-selection approach by Agrobacterium tumefaciens-mediated transformation. The nutritional composition of two lines of transgenic rice plants (RTB5 and RTB11) was compared with that of its non-transgenic counterpart. The results showed that, except for small differences in dietary fiber and some minerals, there was no significant difference between transgenic rice and conventional counterpart variety with respect to their nutrient composition. Most of measured levels of nutrients were within the range of values reported for other commercial cultivars, showing substantial equivalency. Therefore, the insertion of transgenes did not affect the nutritional composition of transgenic RTB5 and RTB11 rice grains.