• Fisheries and Aquatic Sciences
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• 제9권4호
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• pp.135-139
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• 2006

We optimized the biological and physical parameters for DNA delivery into thalli of the red alga Porphyra yezoensis using a particle bombardment device. The efficiency of transformation was determined using the ${\beta}-glucuronidase$ (GUS) assay. The optimal helium pressure, distance of tungsten particle flight, and ratio of DNA to tungsten particles were $23kgf/cm^2$, 8 cm, and $5{\mu}g/mg$ tungsten, respectively. During bombardment, osmotic treatment with a mixture of 0.6 M mannitol and sorbitol increased the efficiency of GUS transformation. After 2 days, the blue color indicating GUS activity was observed using a histochemical assay.

• 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 Microbiology and Biotechnology
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• 제15권4호
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• pp.683-688
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• 2005

A transformation system mediated by Agrobacterium tumefaciens is routinely used for the genetic engineering of plants. Here, we report an efficient and stable method for transformation of heterogeneous genes into an industrial Cephalosporium acremonium by using a similar transformation system established in plants. Both the phleomycin-resistant gene and vgb gene were used as screening markers to confirm the success of transformation by either Southern hybridization or PCR amplification. It was found that acetosyringone (AS) was necessary only for protoplast transformation and the heterogeneous genes transferred were integrated into the genome of C. acremonium. The transformation efficiency obtained with this system was much higher than the conventional techniques used for transformation of C. acremonium.

• International Journal of Computer Science & Network Security
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• 제21권12호
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• pp.349-355
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• 2021

The article investigates the conceptual principles of transformation of industrial parks into eco-industrial ones in the conditions of sustainable development. It is substantiated that the concept of sustainable development in the transformation of industrial parks is to grow industry and jobs, modernize production and introduce innovative technologies, resource and energy efficiency, reduce greenhouse gas emissions and waste storage, social protection of local communities and create favorable infrastructure. It is determined that for the transformation of industrial parks, it is necessary to improve regulatory changes, introduce criteria for compliance of industrial parks and the importance of their consideration, ensure park management by the management company and create favorable incentives for industrial entry into industrial symbiosis. It is proved that industrial parks can be an incentive for industrial development and competitiveness of enterprises. The availability of talented human capital, attractive territories, minerals, energy and mineral resources, developed domestic market, agricultural potential, transport networks is becoming an attractive place for investment and development. Industrial parks need investment. Transformation into eco-industrial parks through the implementation of sustainable development goals opens additional opportunities for access to investment funds and contributes to the implementation of growth and prosperity strategies of the country.

• 한국육종학회지
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• 제41권3호
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• pp.252-260
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• 2009

1. 본 연구에서는 공동배양 배지에 Agrobacterium 성장 억제물질인 silver nitrate를 첨가하고 변온과 여과지처리를 추가하여 공동배양 기간을 7일로 늘였으며, 또한 항산화 물질 3종을 공동배양 배지에 첨가하여 세포의 oxidative burst를 최소화함으로써 벼 형질전환효율을 높일 수 있었다. 또한 이 방법을 적용하여 형질전환이 어려운 품종을 대상으로도 형질전환 식물체를 작성할 수 있었다. 2. 벼 형질전환체의 70%에서 도입유전자 수가 1copy인 것으로 나타나, 적은 수의 유전자가 안정적으로 도입됨을 확인 하였다. 3. 이러한 결과를 바탕으로, 새로운 공동배양 방법을 사용하여 우수한 농업적 형질을 가진 벼 육종 소재 및 품종을 신속하게 개발할 수 있을 것으로 기대된다.

• Journal of Plant Biotechnology
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• 제4권4호
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• pp.135-141
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• 2002

Agrobacterium-mediated transformation has been unsuccessful for monocot plants except for a few important crops such as barley, rice, maize and wheat. We discussed here that a successful transformation of monocots demands certain critical conditions. The requirements for an efficient transformation are a selection of target tissues competent for plant regeneration and Agrobacterium-infection, and various factors promoting Agrobacterium-infection. The factors were divided into two to activate Agrobacterium and to increase plant cell's susceptibility against Agrobacterium. Optimization of these factors significantly increased transformation efficiency of zoysia grass and rice plants. A technical improvement in transformation system for monocots will promote improvement of the breed as well as a study of gene functions in monocots.

• Mycobiology
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• 제45권2호
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• pp.84-89
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• 2017

Fungi of the Metarhizium genus are a very versatile model for understanding pathogenicity in insects and their symbiotic relationship with plants. To establish a co-transformation system for the transformation of multiple M. robertsii genes using Agrobacterium tumefaciens, we evaluated whether the antibiotic nourseothricin has the same marker selection efficiency as phosphinothricin using separate vectors. Subsequently, in the two vectors containing the nourseothricin and phosphinothricin resistance cassettes were inserted eGFP and mCherry expression cassettes, respectively. These new vectors were then introduced independently into A. tumefaciens and used to transform M. robertsii either in independent events or in one single co-transformation event using an equimolar mixture of A. tumefaciens cultures. The number of transformants obtained by co-transformation was similar to that obtained by the individual transformation events. This method provides an additional strategy for the simultaneous insertion of multiple genes into M. robertsii.

• Journal of Forest and Environmental Science
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• 제25권3호
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• pp.195-204
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• 2009

Agrobacterium-mediated genetic transformation has been widely used for the production of genetically modified transgenic plants to obtain specific desired traits. Most of the molecular mechanisms that underlie the transformation steps have been well elucidated over the years. However, a few steps, such as nuclear targeting, T-DNA integration, and Agrobacterium-plant proteins involved remain largely obscure and are still under extensive studies. This review describes the major steps involved in the molecular mechanism of Agrobacterium-mediated transformation and provides insight in the recent developments in studies on the Agrobacterium-mediated genetic transformation system. Some factors affecting the transformation efficiency are also briefly discussed.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2000년도 추계학술발표대회
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• pp.85-91
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• 2000

Aspergillus oryzae is a filamentous fungus classified in the group Aspergillaceae Ascomycetes. It is an important microorganism for industrial production of enzymes and fermented food productions. It secrets large quantities of proteins or enzymes into the culture medium which makes this organism appealing for the production of heterologous proteins. Recently Electric field-mediated transformation method, electroporation, has been applied to fungal transformation. In this study, fungal transformation was carried out by bypassing the protoplast isolation step, decreasing the culturing time and non-protoplast transformation for the increment of transformation efficiency. Transformants were obtained with electroporation in optimal condition 2,500 voltage, 1,540 ohm and 0.50 capacitance. More than 1,000 transform ants were obtained with 6-10 hrs cultured mycelia without enzyme treatment, called non-protoplast transformation.

• Journal of Microbiology and Biotechnology
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• 제9권5호
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• pp.687-690
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• 1999

Bacillus stearothermophilus No. 236 isolated from the soil is a strong xylan degrader producing all the xylanolytic enzymes. However, the strain was discovered to be highly intractable to its transformation. In the present study, we have developed a reliable method for transformation of B. stearothermophilus No. 236 by a systematic examination of several factors which might have an influence on the efficiency of electrotransformation. Notably, we found that the most critical factor influencing the transformation efficiency (TE) was the incubation temperature after pulsing, with its optimum incubation of $37^{\circ}C.\; At\; 50^{\circ}C$, the optimum growth temperature of the B. stearothermophilus strain, the transformants could not be obtained at a recognizable level. The combination of field strength of 7.5 kV/cm along with pulse duration of 10 msec (resistance of $400{\Omega}\; and\; capacitance\; of\; 25{\mu}F$) was shown to be the best electrical parameters at the incubation temperature of $37^{\circ}$. A higher TE was obtained when the cells were harvested at an early-exponential phase. Twenty percent of PEG-8000 in a suspension buffer and an addition of 0.1% glycine in the growth medium resulted in about 4-fold and 3-fold increases in TE, respectively. We also found that the plasmid DNA which had been cycled through the host B. stearothermophilus cells enhanced TE by one order of magnitude higher. Under the presently described conditions, $2.5{\times}10^{5} transformants per ${\mu}g$ DNA was attained.