• 한국균학회지
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• 제20권2호
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• pp.109-117
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• 1992

운동성을 띠는 5가지 부생균과 1종류의 식물병원세균으로 Agrobacterium radiobacter, Bacillus subtilis, B. polymyxa, Pseudomonas aeruginosa, P. fluorescens 및 Xanthomonas malvacearum을 공시하여 이들 세균이 Cochliobolus sativus 분생포자, Fusarium oxysporum f. sp. ciceri의 후막포자, Macrophomina phaseolina의 균핵 및 Phytophthora drechsleri f. sp. cajani 난포자의 분비물에 미치는 화학주성을 서로 다른 무생물적 조건하에서 실험하였다. 균의 분비물의 농도와 세균세포의 끌리는 유인사이에는 정상관$(r=0.76{\sim}0.89)$이 있었다. 유사한 결과로서 화학주성반응과 배양간격에서도 통계적인 정상관이 있다$(r=0.82{\sim}0.95)$. 세균의 화학주성반응은 온도와 pH에 크게 영향을 받는다. 최적 화학주성을 나타내는 온도로서 A. radiobacter는 $25^{\circ}C$, B. polymyxa, P. aerugionosa, p. fluorescens and X. madracearum은 $30^{\circ}C$이었고, B. subtilis는 $35^{\circ}C$가 최적이었다. 최대 화학주성을 나타내는 분비물의 주성은 pH7이고 pH3과 11에서는 buffer와 같았다. 시험용 모세관에 유인물질이나 buffer 중 어느 종류가 채워져도 유인물질이 있는 시험관에 연결되면 세균은 주성이 나타난다는 것이 증명되었다.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.390-394
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• 2004

The genetically modified glyphosate-tolerant soybean contains the following introduced DNA sequences: the EPSPS (5-enol-pyruvylshikimate-3-phosphate synthase) gene from Agrobacterium sp. strain CP4, the 35S promoter from the cauliflower mosaic virus, and the NOS terminator from Agrobacterium tumefaciens. In the present study, detection of these introduced DNAs was performed by amplification using the polymerase chain reaction (PCR). A multiplex PCR method was also applied to prevent false positive results. When primers for 35S promoter, nos3', CTP(chloroplast transit peptide), and CP4 EPSPS (EPSPS from Agrobacterium sp. CP4) were used, positive results were obtained in PCR reactions using DNA from genetically modified glyphosate-tolerant soybeans. There were no false positive results when using DNA from non-genetically modified soybeans. The CP4 EPSPS gene was detected when less than 125 pg glyphosate-tolerant soybean DNA was amplified. Lectin Lel and psb A were amplified from both non-genetically modified and genetically modified glyphosate-tolerant soybean DNA. Multiplex PCR was performed using different primer sets for actin Sacl, 35S promoter and CP4 EPSPS. The actin gene was detectable in both non-genetically modified and glyphosate-tolerant soybeans as a constant endogenous gene. Target DNAs for the 35S promoter, and CP4 EPSPS were detected in samples containing 0.01-0.1% glyphosate-tolerant soybean, although there were variations depending on primers by multiplex PCR. Soybean seeds from five plants of non-genetically modified soybean were co-cultivated for six months with those of genetically modified soybean, and they were analyzed by PCR. As a result, they were not positive for 35S promoter, nos3' or CP4 EPSPS. Therefore, these results suggest there was no natural crossing of genes between glyphosate-tolerant and non-genetically modified soybean during co-cultivation, which indicates that gene transfer between these plants is unlikely to occur in nature.

• 한국토양비료학회지
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• 제20권4호
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• pp.359-367
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• 1987

수종(數種)의 유기산(有機酸), 당류(糖類)(단당(單糖), 복합당(複合糖)) 및 수도근계분비물질(水稻根系分泌物質)에 대(對)한 협생질소고정균(協生窒素固定菌)의 화학주화성(化學走化性)을 검토(檢討)하기 위하여 우리나라 서해간척지(西海干拓地)의 잡초(雜草) 및 수도(水稻) 근조직(根組織)에서 분리동정(分離同定)한 균(菌)을 공시(供試)하여 시험(試驗)하였다. 공시(供試)한 협생질소고정균중(協生窒素固定菌中) Azospirillum lipoferum Ecc 3-1은 유인물질(誘引物質)이 있는 방향(方向)으로 이동(移動)하기 위한 천이환(遷移環)을 만들었으나 Agrobacterium radiobacter Ecc 1-1과 Pseudomonas sp Ecc 4-1은 천이환(遷移環)을 만들지 않했다. 당류(糖類) 및 유기산(有機酸)에 대(對)한 공시세균(供試細菌)의 화학주화성(化學走化性)은 D(+)-glucose나 D(+)-fructose 보다 malic acid나 citric acid에 대(對)한 주화성(走化性)이 높았다. 특(特)히 복합당(複合糖)인 D-galactouronic acid와 L-aspartate는 공시(供試)한 타종(他種)의 당류(糖類)나 유기산(有機酸)보다 가장 높은 주화력(走化力)을 보였다. 수도근분비물질(水稻根分泌物質)에 대(對)한 공시협생질소고정균(供試協生窒素固定菌)의 주화성(走化性)을 수도품종(水稻品種)과 세균간(細菌間)에 서로 상이(相異)한 교호관계(交互關係)를 보였다.

• 한국해양바이오학회지
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• 제16권1호
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• pp.45-54
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• 2024

Microalgae have great potential in the biomedical and pharmaceutical industries as a new type of bioreactor that can produce proteins for specific purposes, including recombinant proteins, pharmaceuticals, and industrial enzymes. Despite the production advantages and importance of microalgae-based expression systems, studies on secretion efficiency are limited. In this study, for stable expression and efficient secretion of the heterologous protein (human SCF and human INFγ) in Chlorella vulgaris, we constructed SP:hSCF:His and SP:hINFγ:His plant binary vectors using the signal peptide (SP) of Chlamydomonas reinhardtii, and we obtained stable transformants through the effective agrobacterium-mediated transformation of these vectors. Transformants with accurately inserted hSCF and hINFγ demonstrated stably increased mRNA and protein expression using RT-PCR and western blotting under the same culture conditions. Following the analysis of the proteins secreted into the culture medium using ELISA, it was confirmed that hINFγ was effectively produced in the transformed C. vulgaris culture medium. The overall findings indicate that the combination of heterologous protein and SP may be crucial for ensuring the expression and secretion of recombinant proteins in Chlorella culture systems.

• Journal of Life Science
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• 제9권1호
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• pp.69-80
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• 1999

Some trials to alter the structure of extracellular polysaccharides by means of biotransformation and microbial modification have been reported. Seaweed alginate was acetylated by intact and resting cells of Pseudomonas syringae ATCC 19304. Glucose analogs such as 3-O-methyl-D-glucose used as sole carbon sources was directly incorporated into curdlan by agrobacterium sp. ATCC 31749. The 2-amino-2-deoxy-D-glucose (glucosamine)and 2-acetamido-2-deoxy-D-glucose (N-acetylglucosamine) were incorporated into microbial cellulose by Acetobacter xylinum ATCC 10245. The changed monomeric composition in pullulan by Aureobasidium pullulans ATCC 42023 as well as zooglan by Zoogoea ramigera ATCC 25935 was another effect of glucose analogs used a carbon source. There was no effect of glucose analogs found in polysacharide-7 (PS-7) produced by Beijerinckia indica. ATCC 21423.

• 생명과학회지
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• 제16권6호
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• pp.938-941
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• 2006

Continuous fermentation process for the production of soluble glucan using mutant Agrobacterium sp. ATCC31750 has been developed in this research. When the concentration of soluble glucan was higher than 6 g/l, the viscosity of the fermented broth was too high and it needs complex separation process to separate from culture broth. Mathematical models which describe the cell growth and glucan production was developed and they kinetic parameters were estimated with experimental data. They are used for the optimization of continuous fermentation process and calculate optimal dilution rate for easy separation of glucan 4 g/l. With continuous fermentation, glucan production rate was increased 1.8 times more than that with batch fermentation.

• Plant Biotechnology Reports
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• 제5권3호
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• pp.245-254
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• 2011

This study describes an efficient protocol for Agrobacterium tumefaciens-mediated transformation of two subgroups of genotype AAA bananas (Musa acuminata cv. Pei Chiao and Musa acuminata cv. Gros Michel). Instead of using suspension cells, cauliflower-like bud clumps, also known as multiple bud clumps (MBC), were induced from sucker buds on MS medium containing $N^6$-Benzylaminopurine (BA), Thidiazuron (TDZ), and Paclobutrazol (PP333). Bud slices were co-cultivated with A. tumefaciens C58C1 or EHA105 that carry a plasmid containing Arabidopsis root-type ferredoxin gene (Atfd3) and a plant ferredoxin-like protein (pflp) gene, respectively. These two strains showed differences in transformation efficiency. The EHA105 strain was more sensitive in Pei Chiao, 51.3% bud slices were pflp-transformed, and 12.6% slices were Atfd3-transformed. Gros Michel was susceptible to C58C1 and the transformation efficiency is 4.4% for pflp and 13.1% for Atfd3. Additionally, gene integration of the putative pflp was confirmed by Southern blot. Resulting from the pathogen inoculation assay, we found that the pflp transgenic banana exhibited resistance to Fusarium oxysporum f. sp. cubense tropical race 4. This protocol is highly advantageous to banana cultivars that have difficulties in setting up suspension cultures for the purpose of quality improvement through genetic transformation. In addition, this protocol would save at least 6 months in obtaining explants for transformation and reduce labor for weekly subculture in embryogenic cell suspension culture systems.

• 한국식품과학회지
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• 제36권3호
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• pp.369-374
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• 2004

유전자재조합 콩 Roundup Ready는 Agrobacterium sp. CP4에서 유래한 유전자를 함유하고 있으며 이 삽입 유전자에서 5-enolpyruvylshikimate-3-phosphate synthase(CP4EPSPS)가 발현된다. 본 연구실에서는 이 단백질을 유전자재조합 콩과 시중에서 구입한 두부 시료에서 CP4EPSPS 특이 단클론 및 다클론 항체를 이용하여 immunoblotting법으로 검사하였다. 분리 정제된 CP4EPSPS 재조합 단백질은 단클론 또는 다클론 항체를 이용할 때 각각 $0.006{\mu}g$ 또는 $0.0006{\mu}g$의 검증 한계치로 확인할 수 있었다. 유전자재조합 콩에 발현된 CP4EPSPS 검증 한계치는 단클론 또는 다클론 항체를 이용할 때 각각 $0.001{\mu}g$과 $0.0001{\mu}g$이었다. 다클론 항체를 이용하여 조사된 9종의 두부에서는 2종에서 양성결과를 확인하였다. 본 연구에서 생산된 단클론 및 다클론 항체를 이용하여 확립된 immunoblotting법은 콩 가공식품 중의 glyphosate 내성 유전자재조합 콩 검사에 적용될 수 있을 것이다.

• Nutrition Research and Practice
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• 제11권1호
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• pp.43-50
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• 2017

BACKGROUND/OBJECTIVES: The present study investigated the hypothesis that a highly pure linear ${\beta}$-1,3-glucan produced by Agrobacterium sp. R259 enhances human natural killer (NK) cell activity and suppresses pro-inflammatory cytokines. SUBJECTS/METHODS: In an eight-week, double-blind, randomized, placebo-controlled clinical trial, 83 healthy adults with white blood cell counts of $4,000-8,000cells/{\mu}L$ were participated and randomly assigned to take two capsules per day containing either 350 mg ${\beta}$-1,3-glucan or placebo. Six participants withdrew their study consent or were excluded due to NK cell activity levels outside the normal range. NK cell activity and serum levels of immunoglobulin G (IgG) and cytokines, such as interferon (IFN)-${\gamma}$, interleukin (IL)-2, IL-4, IL-6, IL-10, IL-12 and tumor necrosis factor (TNF)-${\alpha}$ were measured. RESULTS: NK cell activity and the serum levels of IL-10 were significantly higher from baseline to week 8 in the ${\beta}$-glucan group compared with the placebo group (P = 0.048, P = 0.029). Consumption of ${\beta}$-1,3-glucan also significantly increased NK cell activity compared with placebo after adjusting for smoking and stress status (P = 0.009). In particular, the effect of ${\beta}$-1,3-glucan on NK cell activity was greater in participants with severe stress than in those experiencing mild stress. However, the administration ${\beta}$-1,3-glucan did not significantly modulate the levels of IFN-${\gamma}$, IL-2, IL-4, IL-6, IL-12, TNF-${\alpha}$ and IgG compared with the placebo. CONCLUSION: The results showed that supplementation with bacterial ${\beta}$-1,3-glucan significantly increased NK cell activity without causing any adverse effects. Additionally, the beneficial effect of ${\beta}$-1,3-glucan on NK cell activity was greater in participants experiencing severe stress.

• Journal of Plant Biotechnology
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• 제36권1호
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• pp.75-80
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• 2009

산화스트레스에 내성을 지닌 형질전철 고구마 식물체를 개발하기 위해서 산화스트레스에 의해 발현이 강하게 유도되는 SWPA2 프로모터 또는 CaMV 35S 프로모터에 2-Cys peroxiredoxin (Prx) 유전자가 발현되도록 연결한 형질전철 벡터 (pSP-K, pEP-K)를 제작한 후, 각각 Agrobacterium 매개로 형질전환 하였다. 카나마이신 저항성 배발생 캘러스로부터 체세포배발생 과정을 거쳐 100mg/L kanainycin이 포함된 MS 배지에서 소식물체로 발달하였다. Southern 분석으로 외래 유전자가 안정적으로 고구마 게놈 내로 삽입되었음을 확인하였다. 형질전환 고구마 잎 조직을 대상으로 $20{\mu}M$ methyl viologen에 대한 내성 검정을 조사하여 형질전환 고구마 식물체가 비형질전환 식물체 또는 벡터 대조구 식물체 보다 40% 정도 높은 신화스트레스에 대한 내성을 보여주었다. 선발된 형질전환 식물계는 저온, 건조 등의 여러 기지 환경스트레스 내성검정에 이용될 것이며 향후 복합재해 내성 고구마 계통육성에 이용될 수 있을 것으로 기대된다.