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Dehydroascorbate reductase 과발현 형질전환 감자 식물체의 단백질체 분석
한은희,구영민,김윤희,이신우 한국식물생명공학회 2016 JOURNAL OF PLANT BIOTECHNOLOGY Vol.43 No.2
Ascorbic acid (AsA) is a strong antioxidant/ reducing agent that can be converted to dehydroascorbate (DHA) by oxidation in plants. DHA, a very short-lived chemical, is recycled to AsA by dehydroascorbate reductase (DHAR). Previously, DHAR cDNA was isolated from the hairy roots of the sesame plant, and DHAR-overexpressing transgenic potato plants were generated under the control of the CaMV35S promoter (CaMV35S::DHAR). An increase in transgene expression and ascorbate levels were observed in the transgenic plants. In the present study, proteomic analysis revealed that transgenic plants not only accumulated DHAR in their cells, but also induced several other antioxidant enzyme-related proteins during plant growth. These results suggest that DHAR is important for stress tolerance via induction of antioxidant proteins, and could improve stress tolerance in transgenic potato plants. 아스코르빈산(ascorbic acid, AsA)는 강력한 항산화 물질및 환원제로서 식물에서 산화형 AsA인 dehydroascorbate (DHA)를 활성형인 환원형 AsA로 변환시키는 효소인dehydroascorbate reductase (DHAR)에 의해 생성된다. 선행연구의 결과로서, 본 연구팀은 참깨의 모상근에서 분리한 DHAR 유전자를 이용하여 항시 발현하는 CaMV 35S 프로모터와 괴경 특이적으로 발현하는 patatin 프로모터의 조절하에 과발현 시킨 형질전환 감자를 개발하였다형질전환 감자 식물체들은 비형질전환체 보다 증가된DHAR 활성과 AsA 함량을 보인 바 있다. 본 연구에서는단백질체 분석을 통해서 형질전환 감자 식물체에서DHAR 과발현에 의해 조절되는 단백질들을 조사하였다. 단백질체 분석의 결과로서, 형질전환 감자에서 도입된DHAR 단백질과 다양한 항산화 관련 단백질들이 식물 생장시기 동안 증가 하였다. 본 연구의 결과로서, 도입된유전자인 DHAR이 항산화 단백질들의 발현 증가를 통해형질전환 식물체의 스트레스 내성 기작을 향상 시킬 수있을 것으로 생각된다.
장백도라지의 대량 증식을 위한 조직배양 및 순화 조건 확립
한은희,이신우,손용완,김만배,신용욱,조영손 한국식물생명공학회 2014 식물생명공학회지 Vol.41 No.3
The aim of this study was to establish the conditionof regeneration for white balloon flower (Platycodongrandiflorum DC. cv. Jangback) and to manage for theraising of seedling with in vitro regenerated plants. It wasexamined that 0.5 mg/L of NAA and 1.0 mg/L of BA was thebest composition for the callus and shoot induction (up to600%). NAA was better than IBA for the induction of rootand it took 16.9 days for the induction of rooting on the MSsoild media containing 0.5 mg/L of NAA and the final rootingratio was up to 75%. Out of 5 different bed soils purchasedfrom local market, “Tosil” was identified to be the best forthe acclimation and growth of in vitro regenerated balloonflower. In detail, on 8 weeks after planting of in vitroregenerated plants in pots containing “Tosil” bed soils, theplant hight was increased up to 2-fold (12.8 cm), 3.5-fold(27) for the number of leaf and 1.5-fold (4.5 cm) for the leaflength when compared to the other four bed soils, respectively. Our preliminary results indicate that it is possible to preventthe occurrence of blue balloon flower in the massivecultivated area of white balloon flower by providing theseedlings raised from in vitro regenerated plants.
An efficient transformation method for a potato (Solanum tuberosum L. var. Atlantic)
한은희,구영민,이민경,이신우 한국식물생명공학회 2015 식물생명공학회지 Vol.42 No.2
We found that a long period of in vitro culture is a critical factor on the low transformation rate for a specific potato genotype, Solanum tuberosum L. var. Atlantic when phosphinothricin (PPT) was added to select putative transformants in a solid media. The fresh explants of the newly produced plants from a micro-tuber was able to increase the transformation rate significantly while the old explants prepared from a plant maintained for longer than 6 months in vitro by sub-culturing every 3 ~ 4 weeks resulted in a very low transformation frequency. However, Jowon cultivar was not so much influenced by the period of in vitro culture with high transformation rate (higher than 10.0%). Further research need to be explored for the reason why a particular potato genotype, Atlantic is more vulnerable than the Jowon cultivar during the regeneration stage resulting in the low transformation frequency.
한은희,이혜영,김명신,김용구,한경자,이성은,김희제,김동욱 대한혈액학회 2014 Blood Research Vol.49 No.1
Background The coexistence of t(9;22)(q34;q11.2) and inv(16)(p13q22) chromosomal abnormalities is extremely uncommon, and only a small number of such cases have been reported. Here, we characterized 7 cases of hematologic malignancy exhibiting t(9;22) and inv(16) coexistence. Methods We reviewed the cytogenetic data for hematologic malignancies treated at the Catholic Blood and Marrow Transplantation Center between January 2004 and June 2013. We identified 7 cases exhibiting t(9;22) and inv(16) coexistence. In addition, we analyzed mutations in the IKZF1, NPM1, FLT3, N-RAS, K-RAS, c-KIT, and TP53 genes. Results Four cases of chronic myelogenous leukemia (CML; 1 chronic phase, 2 accelerated phase, and 1 blast phase) and 3 cases of acute myeloid leukemia (AML; 1 de novo and 2 therapy-related) were identified. The percentages of circulating blasts and bone marrow eosinophils were higher in AML cases than in CML cases (53% vs. 5% and 30% vs. 5.5%, respectively). The proportions of each chromosomal abnormality were used along with follow-up karyotyping results to identify secondary changes. In BCR/ABL, a p210 fusion transcript was associated with CML, whereas a p190 fusion transcript was associated with AML. One patient with AML harbored 2 mutations: c-KIT D816V and TP53 E11Q. All patients except 1 with CML blast phase sustained clinical remission after treatment, which included an imatinib mesylate regimen. Conclusion This study shows that observations of bone marrow morphology, initial and follow-up cytogenetic studies, and karyotyping of BCR/ABL1 and CBFB/MYH11 provide valuable information for characterizing hematologic malignancies exhibiting t(9;22) and inv(16) coexistence.
약용작물의 기원 판별에 관한 분자생물학적 기술 개발 현황
한은희,김윤희,이신우 한국식물생명공학회 2015 식물생명공학회지 Vol.42 No.1
Medicinal plants resources are becoming importantassets since their usages have been expanded to the developmentof functional foods for human health, more attractive cosmetics,and pharmaceutical industries. However, their phylogeneticorigins and names are different from each country and quiteoften they are mixed each other resulting in the confusion forconsumers. In particular, when they are very similar basedon their morphological characteristics and distributed asdried roots, it is extremely difficult to differentiate theirorigins even by specialists. Recently, “DNA barcodes” havebeen extensively applied to identify their origin of medicinalplant species. In this review, we tried to overview the currentresearch achievements for the development of suitable“DNA barcodes” regarding to the differentiation of medicinalplant species. Furthermore, more advanced techniques includingamplification refractory mutation system (ARMS)-PCR,multiplex single base extension (MSBE), high-resolution melting(HRM) curve analyses are also discussed for their practicalapplications in the authentification of particular medicinalplant species.