• 제목/요약/키워드: K-L Transformation

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유전자총과 아그로박테리움을 이용한 여러 가지 한국 잔디류의 형질전환체계 확립 (Establishment of a transformation protocol combination particle bombardment with Agrobacterium tumefaciens in different zoysiagrass cultivars)

  • 김종보;김경덕;박대섭
    • 아시안잔디학회지
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    • 제18권3호
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    • pp.141-148
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    • 2004
  • 다양한 Zoysiagrass 4가지 품종들을 식물재료로 사용하여 Agrobacterium만 이용한 방법 그리고 particle bombardment로 배발생캘러스에 상처를 낸 후, Agrobacterium으로 공동배양 시키는 2가지 다른 형질전환 방법을 비교하였다. 예비실험에서 일반적으로 형질전환에 널리 사용되는 kanamycin과 PPT(phospinitricin)의 적적선발농도에 대해서 실험하였는데, kanamycin의 경우 300mg/l 그리고 PPT의 경위 50mg/l의 농도에서 가장 효과적인 선발 효율을 나타내었다. Agrobacterium을 이용한 형질전환은 Agrobacterium을 2일간 배양시킨 다음, 박테리아 농도를 O.D 600nm=1.0-1.2로 맞추고, 배발생캘러스를 30분간 간염 시키는 방법이 효과적이었는데, particle bombardment를 이용하여 캘러스에 상처를 유발시킨 후, Agrobacterium으로 감염시키면 3배 이상 높은 형질전환 수율을 획득할 수 있었다. 이상의 결과는 한국잔디 형질전환에 있어서 particle bombardment과 Agrobacterium을 병행하여 실시한 최초의 보고이고, 이러한 시스템을 기반으로 하여 향후 한국잔디를 포함하여 다른 난지형 및 한지형 잔디의 품종개량에 널리 이용되리라 생각된다.

Comparison of Agrobacterium-mediated of Five Alfalfa (Medicago sativa L.) Cultivars Using the GUS Reporter Gene

  • Lee, Sang-Hoon;Kim, Ki-Yong;Park, Hyung Soo;Cha, Joon-Yung;Lee, Ki-Won
    • 한국초지조사료학회지
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    • 제34권3호
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    • pp.187-192
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    • 2014
  • Alfalfa (Medicago sativa L.) is one of the most important forage legumes in the world. It has been demanded to establish the efficient transformation system in commercial varieties of alfalfa for forage molecular breeding and production of varieties possessing new characteristics. To approach this, genetic transformation techniques have been developed and modified. This work was performed to establish conditions for effective transformation of commercial alfalfa cultivars, Xinjiang Daye, ABT405, Vernal, Wintergreen and Alfagraze. GUS gene was used as a transgene and cotyledon and hypocotyl as a source of explants. Transformation efficiencies differed from 0 to 7.9% among alfalfa cultivars. Highest transformation efficiencies were observed in the cultivar Xinjiang Daye. The integration and expression of the transgenes in the transformed alfalfa plants was confirmed by polymerase chain reaction (PCR) and histochemical GUS assay. These data demonstrate highly efficient Agrobacterium transformation of diverse alfalfa cultivars Xinjiang Daye, which enables routine production of transgenic alfalfa plants.

A New Selection System for Pepper Regeneration by Mannose

  • Kim, Joo-Yean;Min Jung;Kim, Hyo-Soon;Lee, Yun-Hee;Park, Soon-Ho;Lim, Yong-Pyo;Min, Byung-Whan;Yang, Seung-Gyun;Harn, Chee-Hark
    • Journal of Plant Biotechnology
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    • 제4권3호
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    • pp.129-134
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    • 2002
  • We report the development of a new selection system for the transformation of pepper plants by mannose. In order to achieve this, we first tested several factors related to regeneration conditions. Among the 30 inbred lines examined, line P9l5 was able to generate shoots at the highest rate from both cotyledons and hyporotyls in MS media. A dosage curve for optimizing the selection conditions was established by mixing mannose (range 0-50 g/L) and sucrose (range 0-30 g/L). The least selection pressure on shoot formation was created by a mixture of sucrose and mannose at 20 g/L and 15 g/L, respectively, and the threshold for ultimate tissue death was 50 g/L of mannose irrespective of the sucrose concentration. However, we found that mannose itself was not the sole inhibitor of pepper shoot development. High concentrations of sucrose (30 g/L) contributed additively to the inhibition of shoot formation at higher mannose concentrations. Genotype preference is a major factor that enhances regeneration ability in mannose media, as was observed in MS media. P9l5 and P410 line had high regeneration rates under mannose selection conditions in the presence of Agrobacterium infection. Different virulence levels of Agrobacterium strains did change the regeneration rates, probably due to interaction with the specificities of the inbred lines. Taken together, P9l5 offers the best pepper inbred line for transformation and we recommend a selection condition of 20 g/L of sucrose and 15 g/L or more of mannose up to 50 g/L in media.

Agrobacterium-mediated Transformation of Rehmannia glutinosa L. with Resveratrol Gene (RS3) of Peanut

  • Lim, Jung-Dae;Yang, Deok-Chun;Yun, Song-Joong;Chung, Ill-Min;Sung, Eun-Soo;Kim, Myong-Jo;Heo, Kweon;Yu, Chang-Yeon
    • 한국약용작물학회지
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    • 제12권2호
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    • pp.171-178
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    • 2004
  • The objectives of this study were to establish the genetic transformation system of stilbene synthase in Rehmannia glutinosa. Resveratrol, which is both a phytoalexin with antifungal activity and a phytochemical associated with reduced cancer risk and reduced cardiovascular disease, is synthesized in a limited number of plant species including peanut. Resveratrol synthesis is catalyzed by the enzyme stilbene synthase including resveratrol synthase (RS). Stilbene synthase gene (RS3) obtained from peanut, Arachis hypogaea, Fabaceae has been transferred into chinese foxglove, Rehmannia glutinosa by using Agrobacterium mediated transformation. PCR analysis with RS3 primer confirmed that the targeted gene was introduced into the plant genome, 904 bp in size. Further analyses of identification of transformation using developed other molecular techniques and transgenic plants that RS t-DNA introduced to chinese foxglove (R. glutinosa L) and its reaction product, stilbene such as resveratrol will be isolate and characterize using NMR, MS, and HPLC.

국내 육성 감자의 재분화와 형질전환 효율에 미치는 생장조절제의 조성 및 몇 가지 요소 (Effects of Hormone and Several Factors on the Regeneration and Transformation rate of Potato Cultivars Bred in Korea)

  • 이정윤;서효원;조지홍;이신우;윤한대
    • Journal of Plant Biotechnology
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    • 제30권1호
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    • pp.27-33
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    • 2003
  • 국내에서 육종된 감자 품종들에 대하여 재분화 및 형질전환효율에 영향을 미치는 여러 요인들을 조사하고 품종별 최적 조건을 규명하고자 하였다. 최근 육종된 5품종 '추백', '남서', '자심', '조풍'과 '조원'의 잎과 줄기 조직을 재료로 사용하여 배지 내 호르몬 조성별 신초형성률을 조사하였다. 신초형성률은 잎 조직의 경우 '조원'이 GA₃0.1 mg/L, Zeatin 2.0mg/L과 NAA 0.01 mg/L가 포함된 M5 배지에서 90%이상으로 가장 높은 반면 다른 공시품종은 20% 미만이었으며 줄기조직을 치상한 경우 '조풍'이 M5 배지에서 70%의 신초형성률을 나타내었으나. 다른 품종은 25% 이하의 신초형성률을 나타내어 품종별, 조직별로 큰 차이를 나타내었다. 감자의 형질전환 시 Agrobacterium 접종과정에서 75 μM의 acetosyringone을 처리한 경우 잎 조직은 '조원'이 87.9%, 줄기 조직은 '조풍'이 68.4%로 높은 형질전환효율을 나타내어 acetosyringone을 처리하지 않은 대조구에 비하여 1.5∼4.0배의 높은 형질전환효율을 나타냈다.

An efficient transformation method for a potato (Solanum tuberosum L. var. Atlantic)

  • Han, Eun-Hee;Goo, Young-Min;Lee, Min-Kyung;Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • 제42권2호
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    • pp.77-82
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    • 2015
  • 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.

Agrobacterium tumefaciens를 매개로 한 옥수수 유동유전자 Ac 및 Ds에 의한 서양고추냉이 (Armoracia rusticana)의 형질전환 (Transformation of Maize Controlling Element Ac and Ds into Armoracia rusticana via, Agrobacterium tumefaciens)

  • 배창휴;노일섭;임용표;민경수;김동철;김학진;이효연
    • 식물조직배양학회지
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    • 제21권6호
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    • pp.319-326
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    • 1994
  • 십자화과 식물의 유용유전자를 cloning하기 위한 기초연구로서 십자화과 식물인 Armoracia rusicna의 재분화계와 형질전환계를 확립하고, gene tagging을 하기 위하여 binary vector에 삽입된 옥수수의 transposon 유전자 Ac/Ds를 도입한 결과, NAA 0.1 mg/L와 BA 1.0 mg/L를 함유한 MS 배지에서 최적의 shoot를 유기할 수 있었으며, MS 기본배지에 옮기면 쉽게 발근을 유도할 수 있었다. 옥수수의 Ac/Ds의 유전자를 잎에 형질전환시킨 결과 8-10%의 형질전환율을 보였으며, 엽병의 경우에도 4%의 형질전환 식물체가 얻어졌다. Kanamycin 100 mg/L 농도에서 선발한 개체를 PCR 분석 및 Southern blot분석을 행하였던 결과 PCR분석으로부터 Ds유전자가 식물에 도입된 것이 확인되었고, Southern blot 분석으로부터 Ac/Ds 모두가 도입된 것이 확인되었다.

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CENTRAL LIMIT THEOREM ON CHEBYSHEV POLYNOMIALS

  • Ahn, Young-Ho
    • 한국수학교육학회지시리즈B:순수및응용수학
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    • 제21권4호
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    • pp.271-279
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    • 2014
  • Let $T_l$ be a transformation on the interval [-1, 1] defined by Chebyshev polynomial of degree $l(l{\geq}2)$, i.e., $T_l(cos{\theta})=cos(l{\theta})$. In this paper, we consider $T_l$ as a measure preserving transformation on [-1, 1] with an invariant measure $\frac{1}{\sqrt[\pi]{1-x^2}}dx$. We show that If f(x) is a nonconstant step function with finite k-discontinuity points with k < l-1, then it satisfies the Central Limit Theorem. We also give an explicit method how to check whether it satisfies the Central Limit Theorem or not in the cases of general step functions with finite discontinuity points.

Highly efficient production of transgenic Scoparia dulcis L. mediated by Agrobacterium tumefaciens: plant regeneration via shoot organogenesis

  • Aileni, Mahender;Abbagani, Sadanandam;Zhang, Peng
    • Plant Biotechnology Reports
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    • 제5권2호
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    • pp.147-156
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    • 2011
  • Efficient Agrobacterium-mediated genetic transformation of Scoparia dulcis L. was developed using Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pCAMBIA1301 with ${\beta}$-glucuronidase (GUS) (uidA) and hygromycin phosphotransferase (hpt) genes. Two-day precultured leaf segments of in vitro shoot culture were found to be suitable for cocultivation with the Agrobacterium strain, and acetosyringone was able to promote the transformation process. After selection on shoot organogenesis medium with appropriate concentrations of hygromycin and carbenicillin, adventitious shoots were developed on elongation medium by twice subculturing under the same selection scheme. The elongated hygromycin-resistant shoots were subsequently rooted on the MS medium supplemented with $1mg\;l^{-1}$ indole-3-butyric acid and $15mg\;l^{-1}$ hygromycin. Successful transformation was confirmed by PCR analysis using uidA- and hpt-specific primers and monitored by histochemical assay for ${\beta}$-GUS activity during shoot organogenesis. Integration of hpt gene into the genome of transgenic plants was also verified by Southern blot analysis. High transformation efficiency at a rate of 54.6% with an average of $3.9{\pm}0.39$ transgenic plantlets per explant was achieved in the present transformation system. It took only 2-3 months from seed germination to positive transformants transplanted to soil. Therefore, an efficient and fast genetic transformation system was developed for S. dulcis using an Agrobacterium-mediated approach and plant regeneration via shoot organogenesis, which provides a useful platform for future genetic engineering studies in this medicinally important plant.

중환자실 퇴원환자의 집중치료 후 증후군과 삶의 질 (Post-intensive Care Syndrome and Quality of Life in Survivors of Critical Illness)

  • 김수경;강지연
    • 중환자간호학회지
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    • 제9권1호
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    • pp.1-14
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    • 2016
  • Purpose: To investigate the post-intensive care syndrome (PICS) and to analyze the factors affecting the quality of life (QoL) of survivors of critical illness. Methods: Subjects were 114 outpatients who had been discharged from intensive care units of a university hospital in B city, Korea. From July 30 through September 30, 2015, PICS was assessed using the Korean Montreal Cognitive Assessment, Hospital Anxiety-Depression Scale, Korean Instrumental/Activities of Daily Living (K-I/ADL) index, and handwriting transformation, while physical and mental health-related QoL was measured using the SF-12. Results: Of the subjects, 39.5% were screened for mild cognitive disorder and 23.7% experienced handwriting transformation after discharge. Multiple regression analysis revealed that restraint application, current job, time of ${\geq}36$ months after discharge, depression, anxiety, and handwriting transformation accounted for 40.9% of the physical health-related QoL, and depression, anxiety and experience of delirium accounted for 62.4% of the mental health-related QoL. Conclusions: It is necessary to make efforts to reduce restraint application in intensive care units and prevent the occurrence of delirium, with the objective of reducing PICS and improving the QoL of critical illness survivors.

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