• Title/Summary/Keyword: co-transformation

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Magnetic Behaviors of Isolated Fe-Co-Ni Nanoparticles in a Random Arrangement

  • Yang, Choong Jin;Kim, Kyung Soo;Wu, Jianmin
    • Journal of Magnetics
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    • v.6 no.3
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    • pp.94-100
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    • 2001
  • Fe-Co-Ni particles with an average size of 45 and 135 nm are characterized in terms of magnetic phase transformation and magnetic properties at room temperature. BCC structure of Fe-Co-Ni spherical particles can be synthesized from Fe-Co-Ni-Al-Cu precursor films by heating at 600-80$0^{\circ}C$ for the phase separation of Fe-Co rich Fe-Co-Ni particles, followed by a post heating at $600^{\circ}C$ for 5 hours. The average size of nanoparticles was directly determined by the thickness of precursor films. Exchange interactive hysteresis was observed for the nano-composite (Fe-Co-Ni)+(Fe-Ni-Al) films resulting from the short exchange interface between ferromagnetic Fe-Co-Ni particles surrounded by almost papramagnetic Ni-Al-Fe matrix. Arraying the isolated Fe-Co-Ni nano-particles in a random arrangement on $Al_2O_3$substrate the particle assembly showed a behavior of dipole interactive ferromagnetic clusters depending on their volume and inter-particle distance.

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Recent Advances in Pepper Transformation

  • Kim, Hyo-Soon;Lee, Yun-Hee;Lee, Sang-Hee;Park, Yoon-Sik;Choi, Soon-Ho;Min, Byung-Whan;Yang, Seung-Gyun;Harn, Chee-Hark;Jung, Min
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2002.04b
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    • pp.149-149
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    • 2002
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Establishment of a transformation protocol combination particle bombardment with Agrobacterium tumefaciens in different zoysiagrass cultivars (유전자총과 아그로박테리움을 이용한 여러 가지 한국 잔디류의 형질전환체계 확립)

  • Kim Jong-Bo;Kim Kyong-Duck;Park Dae-Sup
    • Asian Journal of Turfgrass Science
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    • v.18 no.3
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    • pp.141-148
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    • 2004
  • In this report, several factors such as infection time and concentration of bacterial suspension, influencing on transient gene expression in Agrobacterium-mediated transformation were evaluated. An appropriate concentration (O.D 600nm = 1.0-1.2) of bateria and 30 min of infection time showed a higher level of GUS expression. To improve transformation efficiency (TE), friable embryogenic calli (FEC) were bombarded by tungsten particles without plasmid DNA, and then co-cultivated with A. tumefaciens LBA4404 which contains pTOK233 super binary vector, carrying neomycin phosphotransferase (NPTII), hygromycin phosphotransferase (hpt) and$\beta-glucuronidase$ (GUS) genes. Three days after co-cultivation with A. tumefaciens and particle bombardment, FEC cultures were transferred to the selection medium (SM: MS medium supplemented with BA 1mg/l, hygromycin 100mg/l, cefotaxime 250 mg/l and vancomycin 200mg/l). They were cultured for 2 weeks and then transferred to the second SM containing hygromycin 50mg/l, cefotaxime 200 mg/l and vancomycin 100mg/l. Later, stable GUS expression was detected 4 to 6 weeks after transfer to the SM. Further, TE from Agrobacterium-mediated transformation after particle bombardment increased to about 3-folds compared with Agrobacterium-mediated transformation without particle bombardment. In the present study, we established an efficient transformation protocol of zoysiagrass by using A. tumefaciens in the combination with particle bombardment for the first time.

Efficient Transformation Method of Soybean Using Meristematic Tissues of Germinating Seeds (발아종자의 분열조직을 이용한 효율적인 콩 형질전환 방법)

  • Kim, Yul-Ho;Park, Hyang-Mi;Choi, Man-Soo;Sohn, Soo-In;Shin, Dong-Bum;Lee, Jang-Yong
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.278-285
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    • 2008
  • An efficient transformation method for soybean [Glycine max (L.) Merr.] using meristematic tissues of germinating seeds has been established. The embryonic axes were excised from germinating seeds of Korean soybean cultivar, Iksannamulkong and 0.5-2 cm long segment containing meristematic tissues were prepared by cutting hypocotyl region. The explants were inoculated with Agrobacterium tumefaciens strain LBA4404 harboring a binary vector with the bar gene as a selectable marker gene and a ${\beta}-glucuronidase$ (GUSINT) reporter gene, and then co-cultured for 7 days on co-cultivation medium (CCM). The meristematic tissues were cultured on shoot induction medium (SIMP6) supplemented with 0.4 mg/l $N_6-benzylaminopurine$ (BAP) and 0.1 mg/l indolebutyric acid (IBA) in the presence of 6 mg/l L-phosphinotricin (PPT) for 2 weeks and the surviving explants were transferred to shoot elongation medium (SEMP6). Transformation was confirmed by Southern blot analysis and the transformation efficiencies ranged from 1.48 to 2.07%. The new modified transformation method was successfully implemented for obtaining several transgenic lines with SMV-CP gene. It is expected that this method could efficiently be used for the transformation of recalcitrant soybean cultivars.

Establishment of an Efficient Agrobacterium Transformation System for Eggplant and Study of a Potential Biotechnologically Useful Promoter

  • Claudiu Magioli;Ana Paula Machado da Rocha;Pinheiro, Marcia-Margis;Martins, Gilberto-Sachetto;Elisabeth Mansur
    • Journal of Plant Biotechnology
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    • v.2 no.1
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    • pp.43-49
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    • 2000
  • An efficient and reliable Agrobacterium transformation procedure based on TDZ (thidiazuron)-induced organogenesis was established and applied to six Brazilian eggp1ant varieties. Optimum transgenic plants recovery was achieved upon the study of the following parameters affecting transformation efficiency, using F-100 variety as a model: i) explant source; ii) pre-culture period; iii) physical state of the pre-culture medium and iv) coculture conditions. The highest frequency of kanamycin-resistant calli derived from leaf explants (5%) was obtained without a pre-culture period and co-cultivation for 24 h in liquid medium followed by five days on solid RM (regeneration medium). For cotyledon explants, best results were achieved upon a pre-culture of 24 h in liquid RM and a co-cultivation period of 24 h in liquid RM followed by three days in solid RM, resulting in a transformation Sequency of 22.7%. Kanamycin-resistant organogenic calli were also obtained from cultivars Emb, Preta Comprida, Round nose Shaded, Campineira and Florida Market. The expression pattern of an epidermis-specific promoter was studied using transformants expressing a chimaeric construct comprised by the promoter Atgrp-5 transcriptionally fused to the coding region of the gus gene. The expression pattern was similar to that previously observed in tobacco and Arabidopsis thaliana, with preferential expression at the epidermis and the stem phloem. These results support the idea that the Atgrp-5 promoter can be used to drive defense genes in these tissues, which are sites of pathogen interaction and spread, in programs for the genetic improvement of eggplant.

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GUS gene expression and plant regeneration via co-culturing with Agrobacterium in grapevine (Vitis vinifera) (Agrobacterium 공동배양을 이용한 포도 재분화율 향상과 GUS 유전자의 발현)

  • Kim, Se-Hee;Kim, Jeong-Hee;Kim, Ki-Ok;Do, Gyeong-Ran;Shin, Il-Sheob;Cho, Kang-Hee;Hwang, Hae-Seong
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.308-314
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    • 2011
  • Efficient transformation and regeneration methods are a priority for successful application of genetic engineering to vegetative propagated plants such as grape. In this study, methods for Agrobacterium tumefaciens-mediated transformation and plant regeneration of grapevine (Vitis vinifera) were evaluated. Tamnara, Heukgoosul, Heukbosek, Rizamat were co-cultivated with Agrobacterium strains, LBA4404 containing the vector pBI121 carrying with CaMV 35S promoter, GUS gene as reporter gene and resistance to kanamycin as selective agent. Seven percent of the maximum regeneration frequency was obtained from co-cultivated with explants from Rizamat with LBA4404 strain on selection medium with kanamycin. The addition of acetosyringone, 200 ${\mu}m$ in virulence induction step was a key factor for successful GUS reporter gene expression in grapevine transformation. Transgenic plants showed resistance to kanamycin and the GUS positive response in leaf ($T_0$) stem ($T_0$) and petiole ($T_0$).

Optimization of Agrobacterium tumefaciens-Mediated Transformation of Xylaria grammica EL000614, an Endolichenic Fungus Producing Grammicin

  • Jeong, Min-Hye;Kim, Jung A.;Kang, Seogchan;Choi, Eu Ddeum;Kim, Youngmin;Lee, Yerim;Jeon, Mi Jin;Yu, Nan Hee;Park, Ae Ran;Kim, Jin-Cheol;Kim, Soonok;Park, Sook-Young
    • Mycobiology
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    • v.49 no.5
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    • pp.491-497
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    • 2021
  • An endolichenic fungus Xylaria grammica EL000614 produces grammicin, a potent nematicidal pyrone derivative that can serve as a new control option for root-knot nematodes. We optimized an Agrobacterium tumefaciens-mediated transformation (ATMT) protocol for X. grammica to support genetic studies. Transformants were successfully generated after co-cultivation of homogenized young mycelia of X. grammica with A. tumefaciens strain AGL-1 carrying a binary vector that contains the bacterial hygromycin B phosphotransferase (hph) gene and the eGFP gene in T-DNA. The resulting transformants were mitotically stable, and PCR analysis showed the integratin of both genes in the genome of transformants. Expression of eGFP was confirmed via fluorescence microscopy. Southern analysis showed that 131 (78.9%) out of 166 transformants contained a single T-DNA insertion. Crucial factors for producing predominantly single T-DNA transformants include 48 h of co-cultivation, pretreatment of A. tumefaciens cells with acetosyringone before co-cultivation, and using freshly prepared mycelia. The established ATMT protocol offers an efficient tool for random insertional mutagenesis and gene transfer in studying the biology and ecology of X. grammica.

The Repeat Heat Treatment Behavior of Double Remelted Fe-Co Ultra-high Strength Steel. - Part. 1 Microstructure Control (Fe-Co기 고인성 고강도강의 반복 열처리 거동 - Part 1. 조직제어)

  • Yoon, Bo-Hee;Park, Kyoung-Tae;Lee, Tae-Hyuk;Kim, Jae-Hoon;Kim, Hong-Kyu;Lee, Seong;Lee, Jong-Hyeon
    • Journal of Korea Foundry Society
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    • v.32 no.1
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    • pp.32-37
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    • 2012
  • In this study, microstructural evaluation was carried out on secondary hardening type ultrahigh strength steel, Fe-Co-Ni composition. This paper as a first part of whole research presented the microstructural behavior by cyclic heat treatment. The cyclic heat treatment method includes normalizing, stress relieving, solution treatment and aging. Especially, solution treatments performed triple times to get maximized solution hardening. Phase transformation and microstructure were observed by using optical microscope (OM), Electron back-scattered diffraction (EBSD) and X-ray stress analyzer. During the triple solution treatment, size of grain boundary was dramatically decreased by generating a packet from the martensite transformation of residual austenite in the inner part of grain, whereas the hardness increase was not significant.