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중금속 오염 토양에서 분리한 니켈 내성 균주의 동정과 유전체 분석을 통한 잠재적 기작 연구
이슬 ( Seul Lee ),( Anamika Khanal ),여준구 ( Junkoo Yeo ),( Kathyleen Nogrado ),조아현 ( Ahyeon Cho ),송윤진 ( Yoonjin Song ),권미지 ( Miji Kwon ),이지훈 ( Ji-hoon Lee ) 한국환경농학회 2018 한국환경농학회 학술대회집 Vol.2018 No.-
The widespread use of metals influenced many researchers to examine the relationship between heavy metal toxicity and bacterial resistance. In this study, we have inoculated heavy metal contaminated soil from Jang-hang region of South Korea in the nickel-containing media (20 mM Ni) for the enrichment. Among dozens of the colonies acquired from the several transfers and serial dilutions with the same concentrations of Ni, 3 colonies with morphological differences were chosen for further studies, which were named as strains Ni-a, Ni-2 and Ni-3. The isolates were identified for their phylogenetic affiliations by using 16S rRNA gene analysis. The strains Ni-2 and Ni-3 were close to Cupriavidus metallidurans and were found to be resistant to antibiotics of vancomycin, erythromycin, chloramphenicol, ampicillin, gentamicin, streptomycin and kanamycin by disk diffusion method. Of the isolated strains, Ni-2 was sequenced for the whole genome, since the Ni-resistance seemed to be better than the other two strains. From the genome sequence we have found that were a total of 89 metal-resistance-related genes including 11 Ni-resistance genes, 41 heavy metal (As, Cd, Zn, Hg, Cu, and Co)-resistance genes, 22 cation efflux genes, 4 metal pumping ATPase genes, and 11 metal transporter genes.
이슬 ( Seul Lee ),( Anamika Khanal ),( Kathyleen Nogrado ),송형근 ( Hyung-geun Song ),조유성 ( Yu-sung Cho ),이지훈 ( Ji-hoon Lee ) 한국환경농학회 2021 한국환경농학회지 Vol.40 No.4
BACKGROUND: Salinity is one of the major limiting factors in agriculture that affect the growth and productivity of crops. It is economically difficult to artificially purify the soil affected by salt. Therefore, the use of plant growth-promoting bacteria (PGPB) in an effort to reduce stress caused by salt is emerging as a cost-effective and environment-friendly method. In this study, the purpose was to isolate the salt-tolerant bacteria from the rhizosphere soil and identify their ability to promote plant growth under salt stress condition. METHODS AND RESULTS: The isolates KST-1, KST-2, AST-3, and AST-4 that showed plant growth-promoting activity for barley in salt conditions were close to Bacillus cereus (KST-1, KST-2, and AST-4) and Bacillus thuringiensis (AST-3) and showed high salt tolerance up to 7% of additional NaCl to the media. When inoculated to barley, the strains had only minor effect on the length of the barley. However, the concentrations of chlorophyll in the barley leaves were found to be higher from the bacteria-inoculated pots than those from the uninoculated control. In particular, the chlorophyll concentration in Bacillus cereus AST-4 experiment was 5.45 times higher than that of the uninoculated control under the same experimental condition. CONCLUSION(S): The isolated salt-tolerant bacteria were found to influence on chlorophyll concentration of the barley. As represented by the strain AST-4, microbes may suggest a cost-effective and environmentally benign method to alleviate salt stress of crops cultivated in salt-accumulated soils such as reclaimed lands.
살코스키 시약과 Indole-3-acetic Acid의 반응 중 트립토판의 간섭
이슬 ( Seul Lee ),어나미까커날 ( Anamika Khanal ),여준구 ( Junkoo Yeo ),노그라도캐시린 ( Kathyleen Nogrado ),조아현 ( Ahyeon Cho ),송윤진 ( Yoonjin Song ),권미지 ( Miji Kwon ),이지훈 ( Ji-hoon Lee ) 한국환경농학회 2018 한국환경농학회 학술대회집 Vol.2018 No.-
Indole-3-acetic acid (IAA) is the most common auxin hormone produced in the plant and is very important because it regulates various aspects of plant growth and development. Tryptophan is the main precursor for IAA while IAA can be synthesized by two pathways i.e., tryptophan-dependent pathway and tryptophan-independent pathway. There have been many studies using Salkowski regent to measure the IAA production by bacteria. In this study, we have found that there was the interference of tryptophan on the colorimetric estimations of indole-3-acetic acid (IAA) by using Salkowski reagent (35% HClO4 and 10 mM FeCl3), which developed a pink color complex of tris-(indole-3-acetate) iron(III) with IAA. Moreover, we detected the effect of different concentration of tryptophan (100, 200, and 300 μM) on different concentration of IAA (100, 200, and 300 μM). The absorbance on IAA for the tested concentrations was found to be increased with the increasing concentration of tryptophan. It seems that both the IAA and tryptophan show the maximum absorbance at the similar wavelengths. It was also observed that the difference between the predicted and measured concentrations of IAA became larger as the predicted IAA concentration decreased. From this study, it is expected that tryptophan would interfere with accurate measurements of IAA, and therefore care should be taken for the measurement of IAA generated by the precursor, tryptophan.
Resveratrol 처리한 HeLa세포에서 angiogenin과 vascular endothelial growth factor 발현유도에 따른 세포이동촉진
조이슬(I-Seul Joe),정신구(Sin-Gu Jeong),조광원(Goang-Won Cho) 한국생명과학회 2014 생명과학회지 Vol.24 No.4
Resveratrol (RSV)은 천연 폴리페놀계 화합물로 세포분열, 성장, 세포이동 등과 같은 다양한 효과가 보고되었다. Angiogenin (ANG)은 Vascular endothelial growth factor (VEGF)와 함께 세포의 증식, 신생혈관형성, tubular structures의 형성, 세포이동 등을 이끄는 중요한 단백질이다. 본 연구에서는 RSV에 의한 세포이동효과를 HeLa 세포에서 관찰하였다. Real-time PCR을 통해 HeLa 세포에 RSV 0~50 μM을 24시간 동안 처리하였을 때, 농도에 따른 ANG, VEGF 유전자 발현이 의미 있게 증가 하였다. 같은 방법으로, RSV 50 μM을 시간에 따라(0~48시간)처리하여 실험하였다. 그 결과, RSV 50 μM을 24시간 동안 처리하였을 때 ANG, VEGF 유전자 발현이 가장 높게 증가하였고, ANG 단백질 분석에서도 동일한 결과를 얻었다. 또한, MTT assay를 이용한 세포 독성연구에서, RSV 50 μM의 농도에서는 영향을 미치지 않음을 관찰하여, 이를 최적의 조건으로 결정하였다. RSV가 처리된 세포에서 세포이동효과를 조사하기 위해 wound-healing assay를 수행하였다. RSV가 처리된 그룹에서 세포이동이 의미 있게 증가하였다. 따라서, 본 연구에는 RSV에 의해 ANG, VEGF의 발현이 증가했고, 이에 따라 세포이동이 향상됨을 관찰하였다. Resveratrol (RSV), a natural polyphenolic compound, is a modulator for cell division and cell migration, and has diverse beneficial properties. Angiogenin (ANG) and vascular endothelial growth factor (VEGF) are considered to be important mechanisms for cell proliferation, angiogenesis, the formation of tubular structures, and migration. In this study, we investigated whether RSV has a migratory effect in HeLa cells. When cells were treated with 0~50 μM of RSV for 24 hr, the expression of ANG and VEGF was significantly increased in a dose dependent manner measured by real-time PCR. Similarly, we performed time dependent experiments for 50 μM RSV treated cells and identified the optimized time at 24 hr. The increased expression in RSV treated cells was confirmed by Western blot analysis. To examine the toxic effects of RSV at the determined conditions, MTT assays were performed. The viabilities were unchanged for 0~50 μM RSV treated cells, while they decreased at 100 μM RSV. To examine the effect of migration in RSV treated cells, we performed a wound-healing assay. The migratory rates were significantly enhanced in the RSV treated group. In this study, we found that RSV induces an increase in the expression of migration factors ANG, VEGF, and enhances cell migration for the determined conditions.
이슬 ( Seul Lee ),김준태 ( Jun-tae Kim ),조순재 ( Soon-jae Cho ),조범연 ( Beom-yeon Cho ),정서훈 ( Seo-hoon Jeong ),김형훈 ( Hyung-hoon Kim ),심현민 ( Hyeon-min Shim ) 한국정보처리학회 2019 한국정보처리학회 학술대회논문집 Vol.26 No.2
유해조수 퇴치 드론의 시스템은 10.525GHz에서 동작하는 Microwave Doppler Radar Sensor를 이용해 경작지에 침입한 조수를 인식한다. 그리고 사용자에게 위험요소에 대한 알림 서비스를 제공한다. 이후 사용자는 드론의 카메라와 카메라 전용 애플리케이션을 이용해 경작지를 실시간으로 보며 드론을 직접 조종하고 Speaker와 Neo Pixel LED Ring를 이용해 유해조수의 시각과 청각을 자극해 도망을 유도하며 퇴치한다. The drone uses a Microwave Doppler Radar Sensor which operates at 10.525GHz to recognize harmful animal which intruded into the arable land. Moreover provide user with notification services on risk factors. Subsequently, the user the drone's camera and a camera-only app to watch the farmland in real-time, steer the drone directly, and use the NeoPixel LED ring and the speaker to stimulate the harmful animal's sight and hearing to induce escape.
인간 골수유래-중간엽 줄기세포(hBM-MSCs)에서 PDE4 억제조절을 통한 신경세포 분화 효율 개선
정다희(Da Hee Jeong),조이슬(I-Seul Joe),조광원(Goang-Won Cho) 한국생명과학회 2016 생명과학회지 Vol.26 No.12
인간 중간엽 줄기세포(hMSCs)는 신경세포(neuron-like cells)를 포함한 다양한 세포로 분화할 수 있는 능력을 지닌 성체 줄기세포(adult stem cells)이다. 본 연구에서는 인간의 골수유래-중간엽기세포(bone marrow-mesenchymal stem cells; hBM-MSCs)를 이용한 신경분화에서 신경세포 표지자(neuronal marker)인 NF-M, Tuj-1 뿐만 아니라 성상세포 표지자(glial marker)인 GFAP의 발현 역시 의미 있게 증가함을 real-time PCR, Western blot, and immunocytochemical staining법을 통하여 관찰하였다. 이를 개선하기 위하여, 신경분화에 중요한 신호전달자(signal intermediator)인 PDE4를 억제한 후 신경분화를 유도하였다. PDE4 억제자인 rolipram 혹은 resveratrol를 각각 처리하여 신경분화한 줄기세포(Roli- or RSV-dMSCs)에서 NF-M, Tuj-1의 발현이 증가하였고 반면, GFAP의 발현은 감소함을 real-time PCR, Western blot, and immunocytochemical staining법을 통하여 관찰하였다. 본 연구를 통하여, PDE4를 조절하며 줄기세포의 신경분화를 개선할 수 있음을 보였다. Human bone marrow mesenchymal stem cells (hBM-MSCs) can differentiate into various cell types including osteoblasts, adipocytes, chondrocytes, and myocytes. Previous studies, including our own, have shown that MSCs can also differentiate into neuron-like cells. However, their rate of neuronal differentiation is not sufficient for application to stem cell therapy, which requires well-defined cell types. For this purpose, we first examined the expression of neuronal lineage markers (GFAP, MAP-2, KCNH1, Nestin, NF-M, and Tuj-1) by real-time PCR, western blot, and immunocytochemical staining. The expressions of the astrocyte marker GFAP and neuronal markers NF-M and Tuj-1 increased in neuronal differentiated MSCs (dMSCs). To improve the neuronal differentiation efficiency, PDE4, an important signaling intermediator in the progression of neuronal differentiation, was modulated using well-known inhibitors such as rolipram or resveratrol and then differentiated into neuronal cells (Roli- or RSV-dMSCs). The expressions of NF-M, Tuj-1 were increased while that of GFAP decreased in Roli- and RSV-dMSCs, which were examined by real-time PCR, western blot, and immunocytochemical staining. From these experiments, we have found that the neuronal differentiation efficiency can be ameliorated by the modulation of PDE4 activity.
실시간 중합효소연쇄반응을 이용하여 논 토양 내 질소순환 관련 기능 유전자의 평가
어나미까커날 ( Anamika Khanal ),이슬 ( Seul Lee ),여준구 ( Junkoo Yeo ),노그라도캐시린 ( Kathyleen Nogrado ),조아현 ( Ahyeon Cho ),송윤진 ( Yoonjin Song ),권미지 ( Miji Kwon ),이지훈 ( Ji-hoon Lee ) 한국환경농학회 2018 한국환경농학회 학술대회집 Vol.2018 No.-
The nitrogen cycle and the microbes responsible for mediating this process have an important role in natural ecosystems, terrestrial habitats and also have a major impact on climate change. In this study, we use molecular techniques such as conventional PCR, clone library construction, sequencing, phylogenetic analysis and real-time PCR to explore microbial communities in a rice paddy soil by detecting and quantifying some of the key functional genes that are involved in the process of nitrogen cycle i.e., nitrogen fixation (nifH), hydrazine synthase (hzsA), nitrous oxide reductase (nosZ), copper-containing (nirK), cytochrome cd1- containing nitrite reductase (nirS), nitrite oxidoreductase (nxrB), and ammonium monoxygenase (amoA). The sequence assessment from the clone library targeting these different genes i.e., nifH, hzsA, nosZ, nirK, nirS, nxrB, and amoA showed a high diversity and dominance of bacterial community. Besides that, Real-time PCR using SYBR green dye and some old and recently developed primers specific for each individual gene revealed the high abundance of nxrB gene and low abundance of hzsA gene. This finding will be very helpful in exploring and exploiting microorganism in terrestrial habitat.