• Membrane and Water Treatment
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• 제4권1호
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• pp.1-9
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• 2013

The process of electromembrane transformation of L-lysine monohydrochlorides into their zwitterionic form in four- and two-chamber electrodialysis apparatus was investigated. The process of transformation at various concentrations of lysine monohydrochloride (0.1-0.6 mol.L-1) was studied and it was established that at the optimum density of current optimal concentrations of lysine hydrochloride during electrodyalisis was in the range of 0.2-0.4 mol.L-1. It was determined that the process of total transformation was accomplished when pH of the lysine solution achieved 10. Changes of concentrations of $Cl^-$ ions and lysine diffused into the neighboring chamber were determined depending on the time. The method developed by us allows adjusting the removal coefficient of $Cl^-$ ions during transformation to a maximal value, the losses of lysine diffused into the next chamber after its return to the technological cycle being less than 1.0 %. The specific energy consumption during the process of transformation in two- and four-chamber electrodialyzers was 0.19 and 0.205 A.h.kg-1 and the current efficiency was 75.9 and 73.1 %, correspondingly. Study of the process of electromembrane transformation allowed obtaining zwitterionic form of L-lysine from L-lysine monohydrochloride with minimal reagent and energy consumption.

• 한국초지조사료학회지
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• 제20권2호
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• pp.103-108
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• 2000

To produce of transgenic orchardgrass, the seed-derived calli of orchardgrass (Dactylis glomerata L.) co-cultivated with Agrobacterium turnefaciens EHAlOl harboring binary vector pIG121-Hm were selected with hygromycin and then transferred onto N6 regeneration medium containing 1 rngl l of NAA, 5 rngl l of kinetin, 250 rngl l of carbenicillin and 50 mg/ l of hygromycin. The efficiency of transformation was differed on cultivars, that is, 'Potomac' appeared 12% of transformation efficiency while 'Amba' did 5.5%. The addition of acetosyringone during co-cultivation was a key to successhl transformation of orchardgrass. Transgene fragments were identified by PCR analysis and the constitutive expression of GUS gene was confirmed by Northern blot analysis. (Key words : Acetosyringone, Agrobacterium tumefaciens, Orchardgrass (Dactylis glomerata L.), Transformation)

• Journal of Microbiology and Biotechnology
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• 제25권12호
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• pp.2116-2124
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• 2015

To understand the effects of physicochemical factors on nitrite transformation by microalgae, a lipid-rich Chlorella with high nitrite tolerance was cultured with 8 mmol/l sodium nitrite as sole nitrogen source under different conditions. The results showed that nitrite transformation was mainly dependent on the metabolic activities of algal cells rather than oxidation of nitrite by dissolved oxygen. Light intensity, temperature, pH, NaHCO₃ concentrations, and initial cell densities had significant effects on the rate of nitrite transformation. Single-factor experiments revealed that the optimum conditions for nitrite transformation were light intensity: 300 μmol/m²/s; temperature: 30℃ pH: 7-8; NaHCO₃ concentration: 2.0 g/l; and initial cell density: 0.15 g/l; and the highest nitrite transformation rate of 1.36 mmol/l/d was achieved. There was a positive correlation between nitrite transformation rate and the growth of Chlorella. The relationship between nitrite transformation rate (mg/l/d) and biomass productivity (g/l/d) could be described by the regression equation y = 61.3x (R² = 0.9665), meaning that 61.3 mg N element was assimilated by 1.0 g dry biomass on average, which indicated that the nitrite transformation is a process of consuming nitrite as nitrogen source by Chlorella. The results demonstrated that the Chlorella suspension was able to assimilate nitrite efficiently, which implied the feasibility of using flue gas for mass production of Chlorella without preliminary removal of NO_X.

• Journal of Plant Biotechnology
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• 제29권2호
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• pp.85-92
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• 2002

Agrobacterium을 이용한 벼의 효율적인 형질전환을 위하여 몇 가지 형질전환 조건을 조사하였다. 그리고 조사된 최적의 형질전환 방법을 이용하여 내한성에 관련된 GPAT 유전자를 식물에 도입하였다. 형질전환 조건은 GUS 발현을 통하여 조사되었다. 본 실험에서는 벼 (Oryza sativa L. cv. Dongjin)의 성숙 종자 유래 캘러스를 3일간 전배양한 후 Agrobacterium을 접종하였다. 접종이 끝난 캘러스는 50mg/L CaCl$_2$, 30mg/L acetosyringone, 2 mg/L 2,4-D, 120 mg/L betaine이 첨가된 공동배양 배지 위에서 암조건으로 10일간 공동배양하여 높은 GUS 발현을 관찰할 수 있었다. 이와 같은 방법으로 GPAT 유전자를 식물에 도입한 결과 54%의 높은 형질전환율을 나타내었다. 형질전환 식물체를 southern 분석한 결과 wild type 식물체에서는 GPAT 유전자가 검출되지 않았으나, 형질전환 식물체에서는 GPAT 유전자가 검출되었다. 또한 GPAT 유전자로 형질전환된 5 계통의 T1 세대에서 hygromycin에 대한 저항성과 감수성의 유전비율이 3 : 1로 분리되었다. 따라서 본 실험의 결과에서 검토된 고빈도의 형질전환 시스템은 단자엽식물의 형질전환에 있어서 모델계로 이용될 수 있으리라 기대된다.

• Journal of applied mathematics & informatics
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• 제34권3_4호
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• pp.309-317
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• 2016

Let V be a Euclidean Jordan algebra with a symmetric cone K. We show that for a Z-transformation L with the additional property L(K) ⊆ K (which we will call ’cone-preserving’), GUS ⇔ strictly copositive on K ⇔ monotone + P. Specializing the result to the Stein transformation S_A(X) := X - AXA^T on the space of real symmetric matrices with the property $S_A(S^n_+){\subseteq}S^n_+$, we deduce that S_A GUS ⇔ I ± A positive definite.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.36-36
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• 2018

Soybean is the important crop in Asian countries as protein source, oil production and animal feed. Improving soybean using genetic transformation is the principal tool in nowadays. Developing herbicide resistant transgenic soybean plants through Agrobacterium-mediated transformation has been worked in many previous studied. However, the transformation efficiency is still low. Many attempts try to find the optimum media condition for plant regeneration after infection. After transformation, the plant regeneration is very important condition to promote growth of transgenic plant. In this study, we optimized a regeneration condition for two Korean soybean cultivar, Dawonkong and Pungsannamulkong using cotyledon, cotyledonary nodes and hypocotyl as explant. The results showed that shoot regeneration of cotyledonary nodes on B5 medium containing 2 mg/L 6-benzylaminopurine showed the highest percentage of regeneration in Dawonkong (75.8%) while Pungsannamulkong presented high number of shoots 2.12 shoots per explant. For transformation condition, co-cultivation in 7 days showed a high number of GUS positive expression. Most of explants can survived under media including 5 mg/L of glufocinate which refers phosphinotricin for 2-week selection. Washing with 400 mg/L of cefotaxime in several times and selection in plant regeneration media with 400 mg/L of cefotaxime can prevent bacteria growth, effectively.

• Journal of Plant Biotechnology
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• 제2권2호
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• pp.89-96
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• 2000

For large DNA fragment transformation in dicots and monocots, BIBAC2 vector system was applied to Arabidopsis thaliana and Oryza sativa L. cv. Jinmi as a model plant, respectively. For Arabidopsis, the Th1 gene in T23L3 BAC clone whose size is about 90 kb was used as the target gene source for transformation. Because T23L3 BAC clone was originally constructed in pBelloBAC11, the target gene was reconstructed into BIBAC2. As the results of reconstruction, 476 colonies were survived in selection medium containing 40 mg/L kanamycin. In colony hybridization analysis, 24 out of 476 colonies exhibited positive signals. In the pulsed-field gel electrophoresis analysis, 11 out of 24 positive clones exhibited the band at the location of 90 kb. In Southern hybridization, positive signal band at the location of 90 kb was observed in all 11 transformants. Using these verified clones, Agrobacterium-mediated transformation was applied to Arabidopsis thaliana th1-201 mutant for genetic complementation test. Twelve thousands T$_1$ seeds were harvested, and antibiotic selection test is being analyzed to verify whether these seeds were transformed. for rice, COR356 that contains 150 kb human genomic DNA in a BIBAC2 vector was used as the target gene. As the results of transformation, 151 out of 210 co-cultivated calli were survived in selection medium containing 5 mg/L hygromycin, and 45 out of 151 survived calli were regenerated into plants. Transformation efficiency was 21.6%. Progeny test using 71 seeds is being analyzed now. These results provide the potential that large DNA fragments can be transferred into both dicots and monocot by Agrobacterium-mediate d transformation system.

• 한국작물학회지
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• 제49권1호
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• pp.61-68
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• 2004

A reproducible transformation system via optimized regeneration media for Korean rice cultivars was established using Agrobacterium tumefeciens LBA4404 (pSBM-PPGN; gusA and bar). Although japonica rice genotypes were easier to produce transgenic plants compared to Tongil type cultivars, transformation efficiencies were not always correlated with regeneration efficiencies of non-transgenic callus on the control medium. Regeneration efficiencies of Donganbyeo, Ilmibyeo, and Manchubyeo were over 50% in non-transgenic control, however, transformation efficiencies were significantly low when only sucrose was added to the media as a carbon source. However, the medium, MSRK5SS-Pr (or MSRK5SM-Pr), that contains $5\textrm{mgL}^{-1}$ kinetin, $0.5\textrm{mgL}^{-1}$ NAA, 2 % sucrose (or maltose), 3% sorbitol, and $500\textrm{mgL}^{-1}$ proline, was the most efficient not only for regeneration of non-transgenic callus but also for regeneration of transgenic callus in the presence of L-phosphinotricin (PPT). Average transformation efficiencies of 16 Korean rice cultivars were significantly enhanced by using the optimized medium from 1.5% to 5.8% in independent callus lines and from 2.9% to 19.4% in tromsgenic plants obained. Approximately 98.9% (876 out of 885) transgenic plants obtained on optimized media showed basta resistance. Stable integration, inheritance and expression of gusA and bar genes were continued by GUS assay and PCR and Southern analysis of the bar gene. With Pst1 digestion of genomic DNA of transgenic plants, one to five copies of T-DNA segment were observed; however, 76% (19 out of 25 transgenic plants) has low copy number of T-DNA. The transformants obtained from one callus line showed the same copy numbers with the same fractionized band patterns.

• Plant Resources
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• 제7권1호
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• pp.39-46
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• 2004

To demonstrate the importance of transformation efficiency in independent event, molecular and cytogenetic analysis were conducted with genomic DNA and chromosome of transgenic plants produced by Agrobacterium tumefeciens LBA4404 (pSBM-PPGN: gusA and bar). Selection ratios of putative transgenic calli were similar in independent experiments, however, transformation efficiencies were critically influenced by the type of regeneration media. MSRK5SS-Pr regeneration mediun, which contains 5 mgL$^{-1}$ kinetin, 2% (w/v) sucrose in combination with 3% (w/v) sorbitol, and 500 mgL$^{-1}$ proline, was efficient to produce transgenic plant of rice from putative transgenic callus in the presence of L-phosphinotricin (PPT). With MSRK5SS-Pr medium, transformation efficincies of Nagdongbyeo were significantly enhanced from 3.7% to 6.3% in independent callus lines arid from 7.3% to 19.7% in plants produced, respectively. Stable integration and expression of bar gene were confirmed by basta herbicide assay, PCR amplification and Southern blotting of bar gene, and fluorescence in situ hybridization (FISH) analysis using pSBM-PPGN as a probe. In Southern blot analysis, diverse band patterns were observed in total 44 transgenic plants regenerated from 20 independent PPT resistant calli showing from one to five copies of T-DNA segments, however, the transformants obtained from one callus line showed the same copy numbers with the same fractionized band patterns.

• Journal of Plant Biotechnology
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• 제34권1호
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• pp.37-45
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• 2007

수박은 형질전환이 매우 어렵고 직접적인 신초 유도방법으로는 안정된 형질전환을 기대할 수 없어서, 다른 여러 작물에서 높은 효율을 보였던 캘러스 유래 신초 유도 방법을 도입하고자 하였다. 수박의 최적 캘러스 유도조건은 자엽 절편체의 경우 2.0 mg/L zeatin과 0.1 mg/L IAA이었으며 뿌리 절편체의 경우 2.0 mg/L BA와 0.1 mg/L 2,4-D이었다. NptII 유전자의 선발 항생제는 kanamycin보다는 paromomycin 빠르고 효과적이었으며, 수박의 절편체에 Agrobacterium을 접종 한 후 paromomycin을 125 mg/L 첨가한 배지에서 선발하였다. pmGFP5-ER vector로 형질전환한 후 캘러스 상태에서 형광현미경을 통해 GFP 유전자의 도입을 확인하였으며, 딱딱한 초록색의 캘러스에서 강한 GFP 발현을 관찰하였고, 자엽유래 캘러스의 경우 WM8에서 9.0%, 뿌리유래 캘러스의 경우 WM6에서 8.3%의 가장 높은 GFP 발현 효율을 보였다. GFP 유전자 도입과 같은 방법으로 WMV-CP 유전자가 있는 pWMV2300 vector로 형질전환한 후 캘러스 상태에서 PCR 및 Southern blot 분석을 한 결과, 두 점의 캘러스에서 WMV-CP 유전자가 도입되었음을 확인하였다. 본 연구를 통하여 확립된 수박의 캘러스 유도 시스템은 안정된 수박 형질전환 방법에 기초 자료로서 이용될 것이다.