• Title/Summary/Keyword: K-L transformation

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Study on CsRCI2D and CsRCI2H for improvement of abiotic stress tolerance in Camelina sativa L.

  • Lim, Hyun-Gyu;Kim, Hyun-Sung;Kim, Jung-Eun;Ahn, Sung-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.196-196
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    • 2017
  • Oilseed crop Camelina (Camelina sativa L.) is a suitable for biodiesel production that has high adaptability under low-nutrient condition like marginal land and requires low-input cost for cultivation. Enhanced abiotic stress tolerance of Camelina is very important for oil production under the wide range of different climate. CsRCI2s (Rare Cold Inducible 2) are related proteins in various abiotic stresses that predicted to localized at plasma membrane (PM) and endoplasmic reticulum (ER). These proteins are consist of eight-family that can be divided into tail (CsRCI2D/E/F/G) and no-tail (CsRCI2A/B/E/H) type of C-terminal. However, it is still less understood the function of C-terminal tail. In this study, CsRCI2D/H genes were cloned through gateway cloning system that used pCB302-3 as destination vector. And we used agrobacterium-mediated transformation system for generation of overexpression (OX) transformants. Overexpression of target gene was confirmed using RT-PCR and segregation ratio on selection media. We analyzed physiological response in media and soil under abiotic stresses using CsRCI2D and CsRCI2H overexpression plant. To compare abiotic stresses tolerance, wild type and CsRCI2D/H OX line seeds were sown on agar plate treated with various NaCl and mannitol concentration for 7 days. In the test of growth rate under abiotic stress on media, CsRCI2H OX line showed similar to NaCl and mannitol stress. In the other hand, CsRCI2D OX line showed to be improved stress tolerance that especially increased in 200mM NaCl but was similar on mannitol media. In greenhouse, WT and CsRCI2D/H OX lines for physiological analysis and productivity under abiotic stresses were treated 100, 150, 200mM NaCl. Then it was measured various parameters such as leaf width and length, plant height, total seed weight, flower number, seed number. CsRCI2H OX line in greenhouse did not show any changes in physiological parameters but CsRCI2D OX line was improved both physiological response and productivity under NaCl stress. Among physiological parameters of CsRCI2D OX line under NaCl stress, leaf length and width were observed shorter than WT but it were slightly longer than WT in 200mM NaCl stress. Furthermore, total seed weight of CsRCI2D OX line under stress displayed to decrease than WT in normal condition, but it was gradually raised with increasing NaCl stress then more than WT relatively. These results suggested CsRCI2D might be contribute to improve abiotic stress tolerance. However, function of CsRCI2H is need to more detail study. In conclusion, overexpression of CsRCI2s family can generate various environmental stress tolerance plant and may improve crop productivity for bio-energy production.

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Blockade of vascular angiogenesis by Aspergillus usamii var. shirousamii-transformed Angelicae Gigantis Radix and Zizyphus jujuba

  • Kang, Sang-Wook;Choi, Jung-Suk;Bae, Ji-Young;Li, Jing;Kim, Dong-Shoo;Kim, Jung-Lye;Shin, Seung-Yong;You, Hyun-Ju;Park, Hyoung-Sook;Ji, Geun-Eog;Kang, Young-Hee
    • Nutrition Research and Practice
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    • v.3 no.1
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    • pp.3-8
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    • 2009
  • The matrix metalloproteinases (MMP) play an important role in tumor invasion, angiogenesis and inflammatory tissue destruction. Increased expression of MMP was observed in benign tissue hyperplasia and in atherosclerotic lesions. Invasive cancer cells utilize MMP to degrade the extracellular matrix and vascular basement membrane during metastasis, where MMP-2 has been implicated in the development and dissemination of malignancies. The present study attempted to examine the antiangiogenic activity of the medicinal herbs of Aspergillus usamii var. shirousamii-transformed Angelicae Gigantis Radix and Zizyphus jujube (tAgR and tZj) with respect to MMP-2 production and endothelial motility in phorbol 12-myristate 13-acetate (PMA)- or VEGF-exposed human umbilical vein endothelial cells (HUVEC). Nontoxic tAgR and tZj substantially suppressed PMA-induced MMP-2 secretion. In addition, $25{\mu}g/mL$ tAgR and tZj prevented vascular endothelial growth factor-stimulated endothelial cell transmigration and tube formation. The results reveal that tAgR and tZj dampened endothelial MMP-2 production leading to endothelial transmigration and tube formation. tAgR and tZj-mediated inhibition of endothelial MMP may boost a therapeutic efficacy during vascular angiogenesis.

A photoelastic study of the stress distribution on canine retraction by segmented TMA T-loop spring (Segmented TMA T-loop spring에 의한 견치 후방이동시의 응력분포에 관한 광탄성법적 분석)

  • Yoon, Young-Jooh;Kim, Kwang-Won;Yu, Pil-Sik
    • The korean journal of orthodontics
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    • v.31 no.2 s.85
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    • pp.199-207
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    • 2001
  • The segmented TMA T-loop spring, used for reciprocal space closure and described by Burstone, was used to achievebodily movement of canine. Photoelastic analysis is a technique for the transformation of internal stress into visible light patterns. The two-dimensional photoelastic stress analysis was performed, and stress distribution was recorded by photography. The purpose of this study was to visualize photoelastically the distribution of forces transmitted to the alveolus and surrounding structures using new segmented TMA T-loop spring for canine retraction. The results were as follows: 1. Decreased activation produced decreased stress of upper 1st. premolar extraction site and increased intrusive stress of upper 1st. molar, regardless of T-loop position. 2. At 5mm activation, More posterior positioning of T-loop Produced an increased stress in upper 1st. premolar extraction site. 3. At 3mm activation, More posterior positioning of T-loop produced an increased stress in upper 1st. premolar extraction site and mesial lower half of upper 1st. molar mesio-buccal root. 4. At 1mm activation, More anterior positioning of T-loop produced an increased stress in upper mesial and blew apex area of upper canine root. 5. 0.25 B/L ratio and 3mm activation produced bodily movement of canine. To summarize, desired tooth movement and anchorage requirement is possible by altering the activation and mesio-distal position of the T-loop spring.

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Impact Assessment on the Change of Thermal Environment, According to the Hydraulic Characteristic Urban Regeneration Stream: Cheonggyecheon Case Study (도심재생하천 내 수리적 특성이 열환경 변화에 미치는 영향 평가: 청계천을 대상으로)

  • Kim, Jeong-Ho;Lee, Ju-Seung;Yoon, Yong-Han
    • Journal of Environmental Policy
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    • v.14 no.2
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    • pp.3-25
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    • 2015
  • Our goal is to verify how changes in water's hydraulic characteristics after urban regeneration stream can affect any possible transformation of its thermal environment. To that end, we analyzed changes in numerous physical characteristics the subject stream along with the meteorological factors and thermal environment affected by it. Cheonggyecheon was selected as our subject as it is a great example of successful urban regeneration stream. As for physical characteristics, we allocated Type I (0.0%) and Type II (20.2%), depending on the green coverage ratio. As for numerical characteristics, at the point of Ba in which the riffle ends, the water temperature fell by $0.2^{\circ}C$ and the flow increased from 0.7m/s to 0.9 m/s with the dissolved oxygen increasing from 0.5mg/L to 0.6mg/L. As for meteorological factors surrounding the subject stream, the temperature dropped from $1.1^{\circ}C$ to $1.4^{\circ}C$ on average and relative humidity increased from 6.6% to 8.7%. Furthermore, there was an irregular change in wind velocity. According to the result of the Wet Bulb Globe Temperature (WBGT), the change in the values of Type I and II inside and on the surface of the subject stream was negligible. The downstream temperature in Type I fell from $0.3^{\circ}C$ to $0.6^{\circ}C$ and by $0.8^{\circ}C$ in Type II. As for vertical cooling effect, the change of water level was 120cm in Type I and 140cm in Type II. As for horizontal cooling effects, the value of Type I was increased from the point of Ba where the riffle ends and the value of Type II was on a steady decline.

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Introduction and Expression of PAP gene using Agrobacterium in Scrophularia buergeriana Miquel (Agrobacterium을 이용한 PAP 유전자의 현삼으로 도입 및 형질발현)

  • Yu, Chang-Yeon;Seong, Eun-Soo;Lim, Jung-Dae;Huang, Shan-Ai;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.2
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    • pp.156-165
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    • 2001
  • Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

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Development and Validation of a Predictive Model for Listeria monocytogenes Scott A as a Function of Temperature, pH, and Commercial Mixture of Potassium Lactate and Sodium Diacetate

  • Abou-Zeid, Khaled A.;Oscar, Thomas P.;Schwarz, Jurgen G.;Hashem, Fawzy M.;Whiting, Richard C.;Yoon, Kisun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.7
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    • pp.718-726
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    • 2009
  • The objective of this study was to develop and validate secondary models that can predict growth parameters of L. monocytogenes Scott A as a function of concentrations (0-3%) of a commercial potassium lactate (PL) and sodium diacetate (SDA) mixture, pH (5.5-7.0), and temperature (4-37DC). A total of 120 growth curves were fitted to the Baranyi primary model that directly estimates lag time (LT) and specific growth rate (SGR). The effects of the variables on L. monocytogenes Scott A growth kinetics were modeled by response surface analysis using quadratic and cubic polynomial models of the natural logarithm transformation of both LT and SGR. Model performance was evaluated with dependent data and independent data using the prediction bias ($B_f$) and accuracy factors ($A_f$) as well as the acceptable prediction zone method [percentage of relative errors (%RE)]. Comparison of predicted versus observed values of SGR indicated that the cubic model fits better than the quadratic model, particularly at 4 and $10^{\circ}C$. The $B_f$and $A_f$for independent SGR were 1.00 and 1.08 for the cubic model and 1.08 and 1.16 for the quadratic model, respectively. For cubic and quadratic models, the %REs for the independent SGR data were 92.6 and 85.7, respectively. Both quadratic and cubic polynomial models for SGR and LT provided acceptable predictions of L. monocytogenes Scott A growth in the matrix of conditions described in the present study. Model performance can be more accurately evaluated with $B_f$and $A_f$and % RE together.

The Improvement of surface activity and Emulsification Activity by Transformation of Lipase Gene in Klebsiella sp. KCL-1, Oil-Degrading Bacterium. (Lipase gene의 도입에 의한 유류분해세균 Klebsiella sp. KCL-1의 표면활성과 유화력 향상)

  • 정수열
    • Journal of Life Science
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    • v.14 no.5
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    • pp.834-839
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    • 2004
  • To improve and oil degrading activity, the lipase gene from Pseudomonase sp. was transformed into Klebsiella sp. KCL-l, an oil degrading bacterium. The selected trasformant was named as a KCL-1/pET-Lip. The surface tension of culture broth of KCL-1/pET-Lip was decreased to 33 dyne/cm from 55 dyne/cm using 4% (v/v) soybean oil as sole carbon source. The surface tension were 44 and 37.5 dyne/cm, to 2% (w/v) glucose and 4% (v/v) kerosene medium, respectively. The emulsification activity of the biosurfactant solution containing lipase of KCL-l/pET-Lip improved better than wild type KCL-l. The soybean oil was most efficient carbon source and substrate for surface activity and emulsification activity of KCL-1/pET-Lip. The expression of lipase was confirmed by SDS-PAGE.

Nucleotide Analysis of Phaffia rhodozyma DNA Fragment That Functions as ARS in Saccharomyces cerevisiae

  • Chung, Hee-Young;Hong, Min-Hee;Chun, Young-Hyun;Bai, Suk;Im, Suhn-Young;Lee, Hwanghee-Blaise;Park, Jong-Chun;Kim, Dong-Ho;Chun, Soon-Bai
    • Journal of Microbiology and Biotechnology
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    • v.8 no.6
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    • pp.650-655
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    • 1998
  • The chromosomal DNA fragment from Phaffia rhodozyma CBS 6938 which is able to autonomously replicate in the yeast Saccharomyces cerevisiae was cloned on an integrative URA3 plasmid. Its minimal fragment exhibiting autonomously replicating activiy in the S. cerevisiae gave a higher frequency transformation efficiency than that found for centromere-based plasmid, and enabled extrachromosoma1ly stable transmission of the plasmids in one copy per yeast cell under non-selective culture condition. The 836-bp DNA element lacked an ORF and did not contain any acceptable match to an ARS core consensus. Sequence analysis, however, displayed a cluster of three hairpin-Ioop-sequences with individual $\triangle {G_{25}}^{\circ}C$ free energy value of -10.0, -17.5, and -17.0 kcal. $mor^{-l}$as well as a 9-bp sequence with two base pair mismatches to the S. cerevisiae/E. coli gyrase-binding site. This 836-bp sequence also included one 7-bp sequence analogous to the core consensus of centromeric DNA element III (CDEIII) of S. cerevisiae, but CDEIII-like 7 bp sequence alone did not give a replicative function in this yeast.

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Plant Regeneration Capacity of Calluses Derived from Mature Seed of Perennial Ryegrass Cultivars (페레니얼 라이그라스의 품종에 따른 성숙종자 유래의 캘러스로부터 식물체 재분화)

  • Lee, Ki-Won;Lee, Joung-Kyong;Kim, Ki-Yong;Ji, Hee-Chung;Park, Hyung-Soo;Kim, Kyung-Hee;Lee, Byung-Hyun;Lee, Sang-Hoon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.29 no.4
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    • pp.285-290
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    • 2009
  • A suitable system for plant regeneration has been established for perennial ryegrass (Lolium perenne L.). In order to investigate the effects of genetic variations of perennial ryegrass in tissue culture response, calli were induced from mature seeds of five cultivars, 'Topgun', 'Accent', 'Renenge GLX', 'Tetrellite', 'Bison' and plant regeneration frequency was compared. Significant differences were observed among the cultivars in both callus induction and plant regeneration. Genotype 'Accent' consistently performed best in the callus formation and plant regeneration. These results can be used useful for molecular breeding of perennial ryegrass through genetic transformation.

A Routine System for Generation of Fertile Transgenic Rice Plants Using Biolistic Method

  • Lee Soo-In;Kim Cha-Young;Lim Chae-Oh;Choi Young-Ju;Kim Ho-Il;Lee Sang-Yeol;Lee Sung-Ho
    • Journal of Plant Biotechnology
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    • v.5 no.3
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    • pp.163-168
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    • 2003
  • A routine system based on particle bombardment of embryogenic callus for recovery of fertile transgenic rice (Oryza sativa L.) plants was developed. Embryogenic callus was established within 2-3 months from calli derived from mature seeds of Korean rice cultivar, Nagdongbyeo. The callus was bombarded with the plasmid pRQ6 containing the $\beta$-glucuronidase gene (gusA) and hygromycin phosphotransferase gene (hph, conferring resistance to hygromycin B), both driven by CaMV 35S promoter. Placement of cells on an osmoticum-containing medium (0.2 M sorbitol and 0.2 M mannitol) 4 hrs prior to and 16 hrs after bombardment resulted in a statistically significant increase with 3.2-fold in transient expression frequency gusA. In five independent experiments, the average frequency of transformation showing GUS activities was $8.86\%$. A large number of morphologically normal, fertile transgenic rice plants were obtained. Integration of foreign gene into the genome of $R_0$ transgenic plants was confirmed by Southern blot analysis. GUS and HPT were detected in $R_1$ progeny and Mendelian segregation of these genes was observed in $R_1$ progeny.