• Title/Summary/Keyword: Agrobacterium method

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Storage of Pollen Biofactory for in vitro Growth and rProtein Synthesis (기내배양과 재조합 단백질 생산을 위한 화분 Biofactory의 저장기술의 개발)

  • 박희성;고재철
    • KSBB Journal
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    • v.19 no.3
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    • pp.174-177
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    • 2004
  • A method of collection and long-term storage of viable lily (Lilium longiflorum) pollen grains were developed for their in vitro growth and transformation in consistency. Petroleum ether, n-heptane, cyclohexane and benzene, as pollen collection medium, were determined less toxic to pollen growth in vitro than others tested. Pollen grains, however, lost their growth activity if stored in these solvents more than a week, So, a serial performance, that is, pollen grain collection in these solvents, air-drying and immediate transfer to low temperature condition was determined desirable for keeping the viability much longer. Pollen grains from this storage showed a successful transformation in vitro with a cDNA encoding tissue plasminogen activator (TPA) protein using Agrobacterium via vacuum infiltration according to western blotting analysis.

Antimicrobial Activity and Characterization of Volatile Flavor EXtracts from Agastache rugosa

  • Song, Jong-Ho;Kim, Min-Ju;Kwon, Kyuk-Dong;Lee, Won-Koo;Park, In-Ho
    • Preventive Nutrition and Food Science
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    • v.4 no.2
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    • pp.97-102
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    • 1999
  • Antimicrobial activity and chemical composition of volatile flavor extracts from Agastache rugosa were investigated. The volatile flavor extracts were obtained from leaves and stems of Agastache rugosa by simultaneous distillation extraction (SDE) method. Antimicrobial activity was investigated by disc diffusion and broth dilution methods against several microorganisms of Bacillus cereus, bacillus megaterium, Bacillus subtilis, Corynebacterium xerosis, Staphylo coccus aureus, Staphylococcus epidermidis, Agrobacterium rhizogenes , Agrobacterium tumefaciences, Enterobacter cloacae, Escherichia coli, Salmonella typhi, Vibrio parahaemolyticus, Candida utilis and Saccharomyces cerevisiae. Volatile flavor extractsfrom leaves have strong antimicrobial activity against C.utilis and S.cerevisiae. When 0.12% volatile flavor extracts from fresh leaves were included in the medium, lag phase of C. utilis was extended 6 hr and that of S.utilis and S.cerevisiae was extended 2hr. Further analyses were performed to elucidatethe effective component of the extracts. The major component of volatile flavor was estragole, a phenolic compound. Minor components were determined to be terpenes , alcohols, acids , esters, ketones and aldethydes.

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Associated Nitrogen Fixation in the Rhizosphere of Rice in Saline and Reclaimed Saline Paddy Soil -IV. Chemotaxis of Associative N2-fixing Bacteria to Single or Combined Sugars and Root Exudates (간척지토양(干拓地土壤)의 수도근권(水稻根圈)에서 협생질소고정(協生窒素固定)에 관(關)한 연구(硏究) -제사보(第四報). 협생질소고정균(協生窒素固定菌)의 단당(單糖) 및 복합당류(複合糖類)와 근분비물질(根分泌物質)에 대(對)한 화학주화성(化學走化性))

  • Lee, Sang-Kyu;Suh, Jang-Sun;Ko, Jae-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.20 no.4
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    • pp.359-367
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    • 1987
  • A method for evaluating bacterial chemotactic responses toward several single of combined sugars and sterile mucilage from the different rice cultivars had been tested. Bacterial genus of Azospirillum, Pseudomonas and Agrobacterium were specially identified from the histosphere of different rice cultivars and graminea grasses in saline and reclaimed saline paddy soil. To evaluate chemotaxis of these strains a modification of Fendrik channel method was used. Under this condition Azospirillum lipoferum Ecc 3-1 reacted stereoisomerically fomulating the single migration ring while Agrobacterium radiobacter Ecc 1-1 and Pseudomonas sp Ecc 4-1 did not. Strains specificities of chemotaxis to the single sugar such as D(+)-glucose and D(+)-fructose were less prominent than malic and citric acid. Chemotactic responses to the combined sugar such as D-galacturonic acid and the L-aspartate were found high attracting reaction than other combined sugars. Chemotaxis of associative $N_2$-fixing bacteria to the root exudates of different rice cultivars were differed among bacterial strains and rice cultivars.

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Agrobacterium-Mediated Transformation on a Plant with Saccharomyces cerevisiae Acid Phosphatse Gene(PHO5) (Agrobacterium을 이용한 Saccharomyces cerevisiae Acid Phosphatse 유전자 (PHO5) 의 식물체로의 도입)

  • Ki yong Kim;Dae yuong Son;Yong Gu Park;Won Il Jung;Jin Ki Jo
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.13 no.3
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    • pp.177-183
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    • 1993
  • This study was conducted to obtain the transformed tobacco plants with S. cerevisiae Acid phosphatase gene(PH05) using Agrobacterium tumefaciens and th confirm plant transformation and gene expression. the results obtained were summarized as follows: APase activity of Saccharomyces cereviase NA 87-11A was remarkably showed up as deep red color when assayed by Tohe and Oshima(1974). PH05 fragment, Apase gene, was obtained from pVC727G and the graphically estimated size was about 1.5kb by agarose gel electrophoresis. The sequencing results of 5'end and 3'end of PH05 using dideoxy chain termination method were coinsided with the full length nucleotide already. pBKJ I vector was constructed by isolation of PH05 fragment from pVC727-1 and pBKSI-1 digesred with Sma I and Xba I. Isolated plasmid from transformed A. tumefaciens with constructed pBKJ I when it was electrophoresed with agarose gel. The dosc of tobacco leaf was cocultivated 재소 transformed Agronacterium tumefaciens. Transformed shoots were selected on kanamtcin-containing MS-n/B medium and they were regenerated. The transgenic tobacco plants were elucidated by isolation of genomic DNA and genomic southern hybridization using ${\alpha}-^{32}P$ labelled PH05 fragments. The PH05 in transformed tobacco plants was expressed in leaf, stem and root, and its APase activity was estimated as deep red color by Tohe method.

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Studies on the microbiological assay method for tabtoxin produced in pseudomonas syringae pv. tabaci (Pseudomonas syringae pv. tabaci가 생산하는 tabtoxin의 미생물학적 검색방법에 관한 연구)

  • 백형석;구재관;전홍기
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.310-315
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    • 1989
  • Tabtoxin produced in Pseudomonas syringae pv. tabace irreversibly inhibits its known physiological target, glutamine synthetase so that causes wildfire disease on leaves of host plant. In this study, we examined a rapid and sensitive microbiological method for tabtoxin assay in several media. In minimal A agar medium nd minimal glucose agar medium, growth inhibition zone of Agrobacterium tumefaciens was larger than that of other indicator strain. However, mostly, growth inhibition zone of indicator strains on the minimal glucose agar medium was smaller than that of on the miniaml A agar medium. In complex agar medium, growth inhibithiton zone was not observed in all the tested indicator strains. Pseudomonas syringae pv. tabaci produced more tabtoxin according to the incubation time. When glutamine was added to the minimal glucose agar medium, growth inhkbition zone of Agrobacterium tumefaciens was reduced.

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Development of genetic transformation method of Korean soybean (국내콩 형질전환 기술개발)

  • Jeon, Eun-Hee;Chung, Young-Soo
    • Journal of Plant Biotechnology
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    • v.36 no.4
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    • pp.344-351
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    • 2009
  • Current status of soybean transformation method in Korera was reviewed with recent publications. Most frequently used method for genetic transformation was Agrobacterium-mediated transformation on cotyledonary node which is most popular method used in foreign country. In addition to this, various methods such as sonicationmediated transformation, in planta transformation, and transformation on meristem tissue of germinating seed, have been tried in Korea, even though their efficiencies on repeatability and stability were relatively low. Based on the promising results developed recently by reviewer, several important considerations for successful soybean transformations were suggested. They are 1) proper genotype screening, 2) targeting transformation on exact point, 3) multiple shoot formation, 4) efficient selection pressure, 5) successful shoot elongation, 6) efficient root formation. These are the basic requirements for stable and highly efficient soybean transformation of Korean soybean.

Development of Marker-free TaGlu-Ax1 Transgenic Rice Harboring a Wheat High-molecular-weight Glutenin Subunit (HMW-GS) Protein (벼에서 밀 고분자 글루테닌 단백질(TaGlu-Ax1) 발현을 통하여 쌀가루 가공적성 증진을 위한 마커프리(marker-free) 형질전환 벼의 개발)

  • Jeong, Namhee;Jeon, Seung-Ho;Kim, Dool-Yi;Lee, Choonseok;Ok, Hyun-Choong;Park, Ki-Do;Hong, Ha-Cheol;Lee, Seung-Sik;Moon, Jung-Kyung;Park, Soo-Kwon
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1121-1129
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    • 2016
  • High-molecular-weight glutenin subunits (HMW-GSs) are extremely important determinants of the functional properties of wheat dough. Transgenic rice plants containing a wheat TaGlu-Ax1 gene encoding a HMG-GS were produced from the Korean wheat cultivar ‘Jokyeong’ and used to enhance the bread-making quality of rice dough using the Agrobacterium-mediated co-transformation method. Two expression cassettes with separate DNA fragments containing only TaGlu-Ax1 and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately into the Agrobacterium tumefaciens EHA105 strain for co-infection. Rice calli were infected with each EHA105 strain harboring TaGlu-Ax1 or HPTII at a 3:1 ratio of TaGlu-Ax1 and HPTII. Among 210 hygromycin-resistant T0 plants, 20 transgenic lines harboring both the TaGlu-Ax1 and HPTII genes in the rice genome were obtained. The integration of the TaGlu-Ax1 gene into the rice genome was reconfirmed by Southern blot analysis. The transcripts and proteins of the wheat TaGlu-Ax1 were stably expressed in rice T1 seeds. Finally, the marker-free plants harboring only the TaGlu-Ax1 gene were successfully screened in the T1 generation. There were no morphological differences between the wild-type and marker-free transgenic plants. The quality of only one HMW-GS (TaGlu-Ax1) was unsuitable for bread making using transgenic rice dough. Greater numbers and combinations of HMW and LMW-GSs and gliadins of wheat are required to further improve the processing qualities of rice dough. TaGlu-Ax1 marker-free transgenic plants could provide good materials to make transgenic rice with improved bread-making qualities.

Development of Marker-free Transgenic Rice for Increasing Bread-making Quality using Wheat High Molecular Weight Glutenin Subunits (HMW-GS) Gene (밀 고분자 글루테닌 유전자를 이용하여 빵 가공적성 증진을 위한 마커 프리 형질전환 벼의 개발)

  • Park, Soo-Kwon;Shin, DongJin;Hwang, Woon-Ha;Oh, Se-Yun;Cho, Jun-Hyun;Han, Sang-Ik;Nam, Min-Hee;Park, Dong-Soo
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1317-1324
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    • 2013
  • High-molecular weight glutenin subunits (HMW-GS) have been shown to play a crucial role in determining the processing properties of the wheat grain. We have produced marker-free transgenic rice plants containing a wheat Glu-1Bx7 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using the Agrobacterium-mediated co-transformation method. The Glu-1Bx7-own promoter was inserted into a binary vector for seed-specific expression of the Glu-1Bx7 gene. Two expression cassettes comprised of separate DNA fragments containing only Glu-1Bx7 and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately to the Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring Glu-1Bx7 or HPTII was infected to rice calli at a 3:1 ratio of Glu-1Bx7 and HPTII, respectively. Then, among 216 hygromycin-resistant $T_0$ plants, we obtained 24 transgenic lines with both Glu-1Bx7 and HPTII genes inserted into the rice genome. We reconfirmed integration of the Glu-1Bx7 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the wheat Glu-1Bx7 were stably expressed in the rice $T_1$ seeds. Finally, the marker-free plants harboring only the Glu-1Bx7 gene were successfully screened at the $T_1$ generation.