• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.208-211
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• 2000

본 연구를 통해 환경 유해성을 평가하기 위한 동물세포를 개발했고 이를 이용한 모니터링 방법 연구와 다양한 독성 물질에 대한 반응성을 확인했다. 개발된 독성 모니터링 동물세포(KFC-A10)는 각 독성 물질에 따라 빛의 발현 양이 증가하는 성향을 가지므로 이번 실험에서 사용된 MMC, BPA, ${\gamma}-ray$에 농도 의존적으로 빛의 양이 증가함을 관찰할 수 있었다. 특히 BPA의 경우는 환경호르몬으로 알려진 바 그 estrogenic 효과를 관찰할 수 있었다.

• Journal of Microbiology
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• 제45권1호
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• pp.11-14
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• 2007

Over the last few years, the endophytic bacterial community associated with citrus has been studied as an important component interacting with Xylella fastidiosa, the causal agent of citrus variegated chlorosis(CVC). This bacterium may also colonize some model plants, such as Catharanthus roseus and Nicotiana clevelandii. In the present study, we compared the endophytic colonization of Citrus sinensis and Catharanthus rose us using the endophytic bacteria Klebsiella pneumoniae. We chose an appropriate strain, K. pneumoniae 342 (Kp342), labeled with the GFP gene. This strain was inoculated onto seedlings of C. sinensis and C. roseus. The isolation frequency was determined one week after the inoculation and the endophytic colonization of K. pneumoniae was observed using fluorescence microscopy. Although the endophytic bacterium was more frequently isolated from C. roseus than from C. sinensis, the colonization profiles for both host plants were similar, suggesting that C. roseus could be used as a model plant to study the interaction between endophytic bacteria and X. fastidiosa.

• Journal of Plant Biotechnology
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• 제37권4호
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• pp.465-471
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• 2010

We assessed whether the cow1 mutant defects are associated with growth of Tos17 and T-DNA insertional rice in blue light (BL). Growth of oscow1 mutants which encoded a member of the YUCCA protein family was retarded in BL. Root to shoot ratios of the mutants were reduced about 2 times lower in the absence of NAA and about 2.5 times lower in the presence of NAA; the shoot growth was not significantly changed by NAA addition. Photosynthetic activity of the mutants was however inhibited in high light. Pigment analysis showed significant difference between wild-type (Chl a:b = 3.02) and mutants (3.84). Carotenoid contents of the mutants were also decreased considerably, implying the involvement of cow1 in pigment formation. These findings lead us to suggest that the growth retardation of oscow1 mutant plants by BL results from the difference of photosynthetic activity in part.

• 한국해양바이오학회지
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• 제11권2호
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• pp.52-61
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• 2019

Over the past few decades, microalgae-based biotechnology conjugated with innovative CRISPR/Cas9-mediated genetic engineering has been attracted much attention for the cost-effective and eco-friendly value-added compounds production. However, the discharge of reproducible living modified organism (LMO) into environmental condition potentially causes serious problem in aquatic environment, and thus it is essential to assess potential environmental risk for human health. Accordingly, in this study, we monitored discharged genetically modified microalgae (GMM) near the research complex which is located in Daejeon, South Korea. After testing samples obtained from 6 points of near streams, several green-colored microalgal colonies were detected under hygromicin-containing agar plate. By identification of selection marker genes, the GMM was not detected from all the samples. For the lab-scale environmental risk assessment of GMM, acute toxicity test using rotifer Brachionus calcyflorus was performed by feeding GMM. After feeding, there was no significant difference in mortality between WT and transformant Chlamydomonas reinhardtii. According to further analysis of horizontal transfer of green fluorescence protein (GFP)-coding gene after 24 h of incubation in synthetic freshwater, we concluded that the GFP-expressed gene not transferred into predator. However, further risk assessments and construction of standard methods including prolonged toxicity test are required for the accurate ecological risk assessment.

• The Plant Pathology Journal
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• 제27권2호
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• pp.138-147
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• 2011

Xylogone ganodermophthora, an ascomycetous fungus, is known to cause yellow rot in the cultivated mushroom Ganoderma lucidum. In this study, we investigated the dissemination of this fungal pathogen in G. lucidum grown in cultivation houses. To determine the role of ascospores produced by X. ganodermophthora in disease development, we constructed a green fluorescent protein-labeled transgenic strain. This X. ganodermophthora strain produced a number of ascomata in the tissues of oak logs on which G. lucidum had been grown and on the mushroom fruit bodies. However, the ascospores released from the ascomata were not able to germinate on water agar or potato dextrose agar. Moreover, less than 0.1% of the ascospores showed green fluorescence, indicating that most ascospores of X. ganodermophthora were not viable. To determine the manner in which X. ganodermophthora disseminates, diseased oak logs were either buried in isolated soil beds as soil-borne inocula or placed around soil beds as air-borne inocula. In addition, culture bottles in which G. lucidum mycelia had been grown were placed on each floor of a five-floor shelf near X. ganodermophthora inocula. One year after cultivation, yellow rot occurred in almost all of the oak logs in the soil beds, including those in beds without soil-borne inocula. In contrast, none of the G. lucidum in the culture bottles was infected, suggesting that dissemination of X. ganodermophthora can occur via the cultivation soil.

• ALGAE
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• 제37권1호
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• pp.75-84
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• 2022

Intercellular nutrient and signal transduction are essential to sustaining multicellular organisms and maximizing the benefits of multicellularity. It has long been believed that red algal intercellular transport of macromolecules is prevented by the protein-rich pit plug within pit-connections, the only physical connection between cells. Fluorescein isothiocyanate-dextran and recombinant green fluorescence protein (rGFP) of various molecular sizes were injected into vegetative cells of Griffithsia monilis using a micromanipulator, and intercellular transport of the fluorescent probes was examined. Pit-connections were found to provide intercellular transport of tracers at rates comparable to plasmodesmata in other organisms. The time necessary for the transport to an adjacent cell was dependent on the molecular size and the direction of the transport. Fluorescent dextran of 3 kDa was transported to adjacent cells in 1-2 h after injection and migrated to all cells of the filament within 24 h, but fluorescent dextran of 10-20 kDa took 24 h to transfer to neighboring cells. The migration occurred faster towards adjacent reproductive cells and to apical cells than basally. Fluorescent tracers above 40 kDa and rGFP was not transported to neighboring cells, but accumulated near the pit plug. Our results suggest that pit-connections are conduit for macromolecules between neighboring cells and that these size-specific conduits allow intercellular communication between the vegetative cells of red algae.

• 대한치과마취과학회지
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• 제14권2호
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• pp.101-106
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• 2014

Background: Remifentanil, an ultra-short-acting mu-opioid receptor agonist, is unique from other opioids because of its esterase-based metabolism, minimal accumulation, and very rapid onset and offset of clinical action. Remifentanil can prevent the inflammatory response and can suppress inducible nitric oxide synthase expression in a septic mouse model. However, the effects of remifentanil on human keratinocyte and autophagy have yet to be fully elucidated during hypoxia-reoxygenation. Here we investigated whether remifentanil confers protective effect against hypoxia-reoxygenation in human keratinocyte and, if so, whether autophagy mediates this effect. Methods: The human keratinocytes were cultured under 1% oxygen tension. The cells were gassed with 94% $N_2$, and 5% $CO_2$ and incubated for 24 h at $37^{\circ}C$. To determine whether the administration of affects human keratinocytes hypoxia-reoxygenation injury, cells were then exposed to various concentrations of remifentanil (0.01, 0.1, 0.5 and 1 ng/ml) for 2 h. After remifentanil treatment, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at $37^{\circ}C$. Control group did not receive remifentanil treatment. Normoxia group did not receive hypoxia and remifentanil treatment for 36 h. 3-MA group was treated 3-methyladenine (3-MA) for 1h before remifentanil treatment. Cell viability was measured using a quantitative colorimetric assay with MTT, showing the mitochondrial activity of living cells. Cells were stained with fluorescence and analyzed with Western blot analysis to find out any relations with activation of autophagy. Results: Prominent accumulation of autophagic specific staining MDC was observed around the nuclei in RPT group HaCaT cells. Similarly, AO staining, red fluorescent spots appeared in RPT group HaCaT cells, while the Normoxia, control and 3-MA groups showed mainly green cytoplasmic fluorescence. We here examined activation of autophagy related protein under H/R-induced cells by Western blotting analysis. Atg5, Beclin-1, LC3-II (microtubule-associated protein 1 light chain 3 form II) and p62 was elevated in RPT group cells. But they were decreased when autophagy was suppressed by 3-MA (Fig. 5). Conclusions: Although the findings of this study are limited to an in vitro interpretation, we suggest that remifentanil may have a beneficial effect in the recovery of wound from hypoxia-reoxygenation injury.

• BMB Reports
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• 제55권2호
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• pp.98-103
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• 2022

Increased mRNA levels of cancer upregulated gene (CUG)2 have been detected in many different tumor tissues using Affymetrix microarray. Oncogenic capability of the CUG2 gene has been further reported. However, the mechanism by which CUG2 overexpression promotes cancer stem cell (CSC)-like phenotypes remains unknown. With recent studies showing that pyruvate kinase muscle 2 (PKM2) is overexpressed in clinical tissues from gastric, lung, and cervical cancer patients, we hypothesized that PKM2 might play an important role in CSC-like phenotypes caused by CUG2 overexpression. The present study revealed that PKM2 protein levels and translocation of PKM2 into the nucleus were enhanced in CUG2-overexpressing lung carcinoma A549 and immortalized bronchial BEAS-2B cells than in control cells. Expression levels of c-Myc, CyclinD1, and PKM2 were increased in CUG2-overexpressing cells than in control cells. Furthermore, EGFR and ERK inhibitors as well as suppression of Yap1 and NEK2 expression reduced PKM2 protein levels. Interestingly, knockdown of β-catenin expression failed to reduce PKM2 protein levels. Furthermore, reduction of PKM2 expression with its siRNA hindered CSC-like phenotypes such as faster wound healing, aggressive transwell migration, and increased size/number of sphere formation. The introduction of mutant S37A PKM2-green fluorescence protein (GFP) into cells without ability to move to the nucleus did not confer CSC-like phenotypes, whereas forced expression of wild-type PKM2 promoted such phenotypes. Overall, CUG2-induced increase in the expression of nuclear PKM2 contributes to CSC-like phenotypes by upregulating c-Myc and CyclinD1 as a co-activator.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.519-526
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• 2023

본 연구는 마우스 대식세포 유래 Raw264.7 세포주에서 황기와 산초 1:1 혼합물(AZM-1:1)의 면역 증진 효능을 탐색하였다. Raw264.7 세포에 100-400 ㎍/mL의 A ZM-1:1 처치는 세포 생존율의 변화 없이 inducible nitric oxide synthase mRNA의 발현 증가와 함께 nitric oxide의 생성을 통계적으로 유의하게 증가시켰다. 더불어 A ZM-1:1은 처치 농도 의존적으로 cyclooxygenase-2 mRNA의 유도와 함께 세포 배양액 중 prostaglandin E₂의 함량을 증가시켰다. 또한, AZM-1:1은 tumor necrosis factor-α, interleukin-1β, interleukin-6 및 monocyte chemoattractant protein-1의 전사를 촉진하였다. Immunoblot 분석을 통하여 AZM-1:1은 mitogen-activated protein kinase의 인산화를 증가시키고, inhibitory-κBα의 인산화를 매개한 분해를 촉진하며, p65의 인산화를 증가시킬 수 있음을 확인하였다. AZM-1:1의 처치는 녹색 형광으로 표지된 대장균 파편의 탐식작용을 촉진하였다. 따라서, 이상의 결과는 A ZM-1:1가 대식세포를 포함한 내재면역을 증진시키는 기능성 식의약 소재가 될 수 있음을 나타낸다.

• 미생물학회지
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• 제52권3호
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• pp.249-253
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• 2016

먹물버섯의 하나인 Coprinellus congregatus는 생활사 동안 여러 종의 laccase 효소를 생성한다. 균사 끝 효소와 버섯시원체 효소 및 sclerotium (균핵) 효소들은 모두 이 균의 분화와 관련되었다. 이핵체 균사를 산성 액체배지(pH 4.0-4.5)에 접종하면 새로운 laccase가 합성되어 분비된다. 이 laccase 유전자의 프로모터의 어느 부분이 산 충격의 신호에 관련된 단백질이 결합하는가 분석하기 위하여 녹색형광단백질(green fluorescent protein, GFP) 유전자를 laccase 프로모터 2.0 kb 다음에 연결하고, 이를 형질전환 벡터인 pBARGEM7-1에 삽입함으로써 발현벡터를 구축하였다. 이 promoter-GFP 조합의 5'-region부터 차례로 제거한 짧은 길이의 이 발현벡터를 먹물버섯 교배형 a1균과 a2균에 형질전환 방법으로 도입시키고 phosphinothricin 저항성으로 형질전환체들을 선발하였다. 선발된 형질전환체 a1 (a1TF)과 a2 (a2TF)를 서로 교배하여 동형접합(homozygotic) 이핵체 형질전환체를 만들었다. 이들을 산성 액체배지에서 36시간 배양하고 균체를 모아 confocal microscope를 사용하여 형광을 분석하였다. Laccase 유전자의 전체 프로모터(2.0 kb)를 가진 발현벡터(F0-GFP)를 도입한 동형접합 형질전환체에서는 형광을 보였으나, 그 보다 짧은 길이(1.29 kb 이하)의 프로모터를 가진 형질전환체에서는 형광이 나타나지 않았다. 이 결과에 근거하여 먹물버섯의 산 충격에 대한 신호를 받는 부위가 laccase 유전자 프로모터의 -2.0 kb ~ -1.29 kb 사이에 있을 것으로 추정한다.