• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.141-141
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• 2017

The ubiquitin-proteasome pathway is the major regulatory mechanism in a number of cellular processes for selective degradation of proteins and involves three steps: (1) ATP dependent activation of ubiquitin by E1 enzyme, (2) transfer of activated ubiquitin to E2 and (3) transfer of ubiquitin to the protein to be degraded by E3 complex. F-box proteins are subunit of SCF complex and involved in specificity for a target substrate to be degraded. F-box proteins regulate many important biological processes such as embryogenesis, floral development, plant growth and development, biotic and abiotic stress, hormonal responses and senescence. However, little is known about the F-box genes in wheat. The draft genome sequence of wheat (IWGSC Reference Sequence v1.0 assembly) used to analysis a genome-wide survey of the F-box gene family in wheat. The Hidden Markov Model (HMM) profiles of F-box (PF00646), F-box-like (PF12937), F-box-like 2 (PF13013), FBA (PF04300), FBA_1 (PF07734), FBA_2 (PF07735), FBA_3 (PF08268) and FBD (PF08387) domains were downloaded from Pfam database were searched against IWGSC Reference Sequence v1.0 assembly. RNA-seq paired-end libraries from different stages of wheat, such as stages of seedling, tillering, booting, day after flowering (DAF) 1, DAF 10, DAF 20, and DAF 30 were conducted and sequenced by Illumina HiSeq2000 for expression analysis of F-box protein genes. Basic analysis including Hisat, HTseq, DEseq, gene ontology analysis and KEGG mapping were conducted for differentially expressed gene analysis and their annotation mappings of DEGs from various stages. About 950 F-box domain proteins identified by Pfam were mapped to wheat reference genome sequence by blastX (e-value < 0.05). Among them, more than 140 putative F-box protein genes were selected by fold changes cut-offs of > 2, significance p-value < 0.01, and FDR<0.01. Expression profiling of selected F-box protein genes were shown by heatmap analysis, and average linkage and squared Euclidean distance of putative 144 F-box protein genes by expression patterns were calculated for clustering analysis. This work may provide valuable and basic information for further investigation of protein degradation mechanism by ubiquitin proteasome system using F-box proteins during wheat development stages.

• 한국미생물·생명공학회지
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• 제33권3호
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• pp.169-177
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• 2005

본 연구는 어업이나 농업 등의 인간활동에 의하여 상당한 영향을 받고 있는 순천만 갈대의 근권에서 이루어지는 정화작용에 있어 미생물의 역할을 조사하는 것에 중점을 두었다. 일반적으로 만은 환경 스트레스로 인한 갑작스런 충격을 감소시키는 완충지역으로서의 역할을 하는 것으로 알려져 있다. 갈대근권에서 이루어지는 정화기능에 있어 미생물의 역할을 밝히기 위한 첫 번째 노력의 일환으로 벤젠, 톨루엔, 또는 자일렌이 단일 탄소 및 에너지원으로 첨가된 농화배양법을 사용하여 BTEX를 분해할 수 있는 두 종류의 그람양성 세균을 순수분리하였다. 다양한 온도조건의 BTEX배지에서 이들 세균을 배양하는 동안 BTEX의 분해율 및 성장률을 주기적으로 조사한 결과 $37^{\circ}C$에서 가장 빠른 기질 분해율을 보였고 $42^{\circ}C$의 고온에서도 두 균 모두 잘 성장하는 것으로 나타났다. 본 연구에서 시험한 수은, 카드뮴, 납, 바륨, 철 대부분의 중금속에 강한 내성뿐만 아니라 ampicillin을 비롯한 시험한 5종류의 항생제 모두에 강한 내성을 나타냈다. 16S rRNA 유전자 서열과 다양한 표현형 및 형태학적 특성을 기초로 한 동정의 결과 BTEX를 분해할 수 있는 두 균주는 $95{\%}$상의 신뢰도로 Microbacterium sp.와 Rhodococcus sp.로 나타났고, 각 균주는 Microbacterium sp. EMB-1과 Rhodococcus sp. EMB-2로 명명하였다. 이러한 결과는 주요한 염습지 식물중의 하나인 갈대의 근권에서 살아가는 이들 균주들이 BTEX와 같은 석유로 오염된 근권 환경의 정화작용에 중요한 역할을 할 수 있음을 제시해주었다.

• 한국환경농학회지
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• 제30권4호
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• pp.424-431
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• 2011

본 연구에서 우리는 광양만의 해수와 표층 퇴적물에서 TBTCl 내성세균의 개체수를 조사하였다. 광양만에서 TBTCl 내성세균의 개체수는 해수에서 $2.5{\times}10^3-3.8{\times}10^3$ cfu/mL 범위였으며, 표층 퇴적물에서 TBTCl 내성세균의 개체수는 $3.2{\times}10^5-9.1{\times}10^5$ cfu/g 범위였다. 광양만에서 TBTCl 내성세균의 종조성은 Vibrio spp. (19.2%)가 가장 높은 우점종으로 나타났고, Bacillus spp. (16.2%), Aeromonas spp. (15.2%), Pseudomonas spp. (13.1%), Klebsiella spp. (11.1%), Alteromonas spp. (9.1%), Pantoea spp. (6.1%), Proteus spp. (3.0%), Listeria spp. (2.0%), unidentified (5.0%)의 순으로 우점하였다. 또한 11개의 대표적인 TBTCl 내성균주는 여러 중금속들(Cd, Cu, Hg 및 Zn)에도 내성을 나타내었다. 이들 중에서 가장 강한 TBTCl 내성을 보이는 T7 균주를 선별하여, API 20NE등을 이용하여 본 균주의 형태학적, 생리학적 및 생화학적 특성들을 조사하였다. T7 균주는 16S rRNA gene 염기서열 분석에 의해 Pantoea 속으로 동정되어 Pantoea sp. T7으로 명명되었다. 또한 본 균주는 $500{\mu}M$의 TBTCl 농도에서도 60시간 배양 후에 정상 균주 성장의 50.7%까지 증식하였다. Pantoea sp. T7의 생물학적 TBTCl 분해활성은 GC-FPD 분석에 의해 측정되었는데, $100{\mu}M$의 TBTCl 농도에서 배양 40시간 후에 TBTCl 제거 효율은 62.7%로 나타났다.

• 동의생리병리학회지
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• 제23권1호
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• pp.127-134
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• 2009

In the present study, we investigated the anti-allergic effect of the water extract of Bopyeoyangyeong-jun(BYJ) to cytokines and transcription. To investigate the biological effect of BYJ, We examined cytotoxicity and inflammatory cytokine secretion with RBL-2H3. We examined tumor necrosis factor-alpha(TNF-$\alpha$), interleukin(IL)-4 secretion from RBL-2H3 cell after pre- treatment with Bopyeoyangyeong-jun of $1\;mg/m{\ell}$, $2\;mg/m{\ell}$. RBL-2H3 cell was stimulated with phorbol 12-myristate 13-acetate(PMA) and calcium ionophore A23187. We observed that Bopyeoyangyeong-jun reduced TNF-$\alpha$, IL-4 secretion and mRNA expression in RBL-2H3 cells. Moreover, the expression of levels of cyclooxygenase (COX)-2 mRNA, nuclear factor-kappa B (NF-${\kappa}B$) (p65) protein, ERK MAPK, and the degradation of level inhibitor kappa B-alpha ($I{\kappa}B-{\alpha}$) were down-regulated by BYJ. Taken together, these results indicate that Bopyeoyangyeong-jun hascontrols TNF-$\alpha$, IL-4 secretion on allergic reaction.

• 동의생리병리학회지
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• 제31권2호
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• pp.105-110
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• 2017

Prolonged exposure to solar ultraviolet A (UVA) radiation has been known to cause premature skin aging (photo-aging). UVA radiation generates ROS thereby induce degenerative changes of skin such as degradation of dermal collagen, elastic fibers. Matrix metalloproteinases (MMPs), the proteolytic enzymes have been implicated as a major player in the development of UVA-induced photo-aging. Many studies have been conducted to block the harmful effects of UV radiation on the skin. Recently, we are interested in the availability of fermented red ginseng (FRG) as natural matrix metalloproteinases inhibitors (MMPIs). The efficacy difference between red ginseng and FRG has been compared. Both RG and FRG have no cytotoxic effects below the concentration of $300{\mu}g/ml$. Human dermal fibroblasts (HDFs) were pretreated with FRG or RG for 24h, followed by irradiation of UVA. Then, we measured the intracellular ROS production and the expression of MMP, $IL-1{\beta}$ at the mRNA level. We also examined the intracellular localization of $NF-{\kappa}B$ and MMP-9 on the FRG or RG treated and UVA-irradiated HDFs. FRG decreased the intracellular ROS production elicited by UVA. In addition, FRG decreased the mRNA expression of MMP-3, MMP-9, and $IL-1{\beta}$ more efficiently than RG. Furthermore, FRG suppressed the nuclear localization of $NF-{\kappa}B$, and the expression of MMP-9. Taken together, our results suggest that FRG is promising agents to prevent UVA-induced photo-aging by suppressing MMP expression and inflammation.

• KSBB Journal
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• 제11권2호
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• pp.165-172
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• 1996

자신의 3차구조를 가진 인체 프로인슐린을 얻기 위하여 Staphylococcal 프로테인 A(SPA)의 신호 랩타이드를 이용하여 대장균내에서 분비 발현을 시도하였다. 분비 발현을 위해 T7 프로모터, SPA 리보좀 바인팅 부위, SPA 신호 랩타이드, 프로인슐린 유전자를 연속적으로 연결하여 분비 벡터를 구성하였다. 이 벡터를 대장균에 넣은후 발현을 유도했으 나 면역적으로 반응하는 인체 프로인슐린은 배양액이나 페리프라스믹 공간에서 거의 존재하지 않았으며 세포 내에도 존재하지 않았다. 그러나 말현 유도시 세포 내에 프로인슐린 RNA가 급격히 증가하였으며 구성한 벡터는 실험실적으로 프로인슐린을 전 사(transcription) , 번역 (translation) 할 수 있었다. 이는 프로인슐린이 번역 후 급히 세포내에서 분 해됨을 의미하며 이로 인해 분비된 프로인슐린을 거의 얻을 수 없게 된 것으로 생각된다. 그러나 프로인 슐린의 세포내 안정성을 위해 말토즈 바인딩 프로테인을 융합짝으로 프로인슐린에 연결한 경우 과량의 분비된 인체 프로인슐린을 검출할 수 있었다.

• Journal of Plant Biotechnology
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• 제2권2호
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• pp.79-82
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• 2000

We have isolated a cDNA encoding a calcium-dependent protein kinase (CDPK) in Nicotiana tabacum, which was designated NtCDPK1. Accumulation of the NtCDPK1 mRNA was stimulated by various stimuli, including phytohormones, CaCl$_2$ wounding, fungal elicitors, chitin and methyl jasmonate. The NtCDPK1 gene encodes a functional Ser/Thr protein kinase of which phosphorylation activity is strongly induced by calcium. By analyzing expression of the NtCDPK1-GFP fusion protein and by immunoblotting with antibody which reacts with NtCDPK1, we found that NtCDPK1 is localized in membrane and nucleus in plant cells. Silencing expression of the NtCDPK1 transgene resulted in marked decrease of lateral root development in the transgenic tobacco plants. Yeast two hybrid screening using NtCDPK1 as a bait identified a tobacco homologue of proteasome regulatory subunit 21D7, designated Nt21D7. The 21D7 mRNA has been shown to be predominantly expressed in proliferating tissues in the cell cycledependent manner in carrot. The recombinant NtCDPK1 protein associated with Nt21D7 in vitro, and could phosphorylate the Nt21D7 protein in vitro in the presence of calcium, suggesting that Nt21D7 protein is a natural substrate of NtCDPK1 in tobacco. These results suggest that NtCDPK1 may regulate tell proliferation processes, such as lateral root formation, by regulating specificity and/or activity of proteasome-mediated protein degradation pathway.

• 한국미생물·생명공학회지
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• 제43권2호
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• pp.158-163
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• 2015

본 연구에서, 우리는 S. mutans Ag I/II 재조합단백질에 의해 유도되어진 염증유발 단백질의 발현에 홍삼 40% 에탄올 추출물의 효과를 알아보고자 하였다. 홍삼에탄올추출물은 Ag I/II 재조합단백질에 의해 유도되어진 염증유발물질들의 mRNA와 단백질의 발현을 억제하였다. 더불어 홍삼에탄올 추출물은 NF-κΒ p65가 핵내로 이용하는 것이 억제하였다. 결론적으로 홍삼 40%에탄올추출물은 NF-κB의 활성에 의해 NO 생성과 iNOS 발현이 조절되어지는 것으로 생각되어지며, 염증유발 관련 유전자들의 낮은 발현을 유도하는 것으로 관찰되어졌다.

• Journal of Microbiology and Biotechnology
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• 제16권11호
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• pp.1728-1733
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• 2006

A Gram-negative, strictly aerobic, nonmotile, and nonspore-forming bacterial strain, designated $T5-12^T$, was isolated from compost and characterized using a polyphasic taxonomical approach. The isolate was positive for catalase and oxidase tests. It could degrade DNA, but was negative for degradation of macromolecules such as casein, collagen, starch, chitin, cellulose, and xylan. The DNA G+C content was 36.0 mol%. The predominant isoprenoid quinone was menaquinone 7 (MK-7). The major fatty acids were $iso-C_{15:0}$ (45.6%), $iso-C_{17:0}$ 3OH (17.2%), and summed feature 4 ($C_{16:0}\;{\omega}7c$ and/or $iso-C_{15:0}$ 2OH, 14.9%). Comparative 16S rRNA gene sequence analysis showed that strain $T5-12^T$ fell within the radiation of the cluster comprising members of the genus Sphingobacterium. Strain $T5-12^T$ exhibited lower than 94% of 16S rRNA gene sequence similarity with respect to the type strains of recognized Sphingobacterium species. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain $T5-12^T$ ($=KCTC\;12578^T=LMG\;23401^T=CCUG\;52467^T$) should be classified in the genus Sphingobacterium as the type strain of a novel species, for which the name Sphingobacterium composti sp. novo is proposed.

• Journal of Microbiology and Biotechnology
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• 제30권4호
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• pp.533-539
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• 2020

A quantitative real-time polymerase chain reaction (QPCR) was applied to estimate biokinetic coefficients of Clostridium cadaveris and Clostridium sporogenes, which utilize protein as carbon source. Experimental data on changes in peptone concentration and 16S rRNA gene copy numbers of C. cadaveris and C. sporogenes were fitted to model. The fourth-order Runge-Kutta approximation with non-linear least squares analysis was employed to solve the ordinary differential equations to estimate biokinetic coefficients. The maximum specific growth rate (μ_max), half-saturation concentration (K_s), growth yield (Y), and decay coefficient (K_d) of C. cadaveris and C.sporogenes were 0.73 ± 0.05 and 1.35 ± 0.32 h^-1, 6.07 ± 1.52 and 5.67 ± 1.53 g/l, 2.25 ± 0.75 × 10¹⁰ and 7.92 ± 3.71 × 10⁹ copies/g, 0.002 ± 0.003 and 0.002 ± 0.001 h^-1, respectively. The theoretical specific growth rate of C. sporogenes always exceeded that of C. cadaveris at peptone concentration higher than 3.62 g/l. When the influent peptone concentration was 5.0 g/l, the concentration of C.cadaveris gradually decreased to the steady value of 2.9 × 10¹⁰ copies/ml at 4 h Hydraulic retention time (HRT), which indicates a 67.1% reduction of the initial population, but the wash out occurred at HRTs of 1.9 and 3.2 h. The 16S rRNA gene copy numbers of C. sporogenes gradually decreased to steady values ranging from 1.1 × 10¹⁰ to 2.9 × 10¹⁰ copies/ml. C. sporogenes species was predicted to wash out at an HRT of 1.6 h.