• Journal of Microbiology and Biotechnology
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• 제6권6호
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• pp.468-473
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• 1996

The genome of tomato spotted wilt virus (TSWV) is composed of three RNA segments, S, M, and L RNA and the 5.0 kb M RNA encodes two glycoproteins Gl, G2 and NSm protein of unknown function. In an effort to investigate the function of the NSm protein, antibody was raised against NSm fusion protein overexpressed in Escherichia coli. This antibody was used to detect the NSm protein by using western blot analysis and electron microscopic observation after immunogold labelling. For the cloning of the NSm gene, total RNA extracted from a TSWV infected plant was used for cDNA synthesis and polymerase chain reaction (PCR) instead of going through time-consuming virus purification. A protein band specifically reacting to the NSm antibody was detected from TSWV inoculated plants. The NSm protein was detected in the cell wall fraction and in pellet from low speed centrifugation when the infected plant tissue was fractionated into 4 fractions. In the immuno-electron microscopic observation, gold particles were found around the plasmodesmata of infected plant tissue. These results suggest that the NSm protein of TSWV plays some role in cell-to-cell movement of this virus.

• 접착 및 계면
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• 제15권2호
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• pp.63-68
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• 2014

주형으로 구형의 폴리스티렌을 사용하고 질소와 탄소원으로 시안아미드를 사용하여 열처리 과정을 거친 후 구형의 할로우 메조포러스 질화탄소 물질을 합성하였다. 이때 할로우 메조포러스 질화탄소 물질을 합성하는 과정에서 실리카와 같은 무기물 주형을 사용하지 않기 때문에 이차적인 실리카 제거 공정이 필요 없고 용매를 전혀 사용하지 않는다. 구형의 폴리스티렌 입자는 약 170 nm 크기였고 그리고 할로우 메조포러스 질화탄소 구형입자의 할로우 직경은 약 82 nm, 벽 두께는 약 13 nm이었다. 또한 할로우 메조포러스 질화탄소 물질의 표면적, 나노세공 크기, 세공부피는 각각 $188m^2g^{-1}$, 3.8 nm, $0.35cm^3g^{-1}$이었다. 한편, 할로우 벽은 흑연구조와 유사한 박막층의 쌓임 구조를 가졌으며 이러한 할로우 메조포러스 질화탄소 물질은 연료전지, 촉매, 광촉매, 전자방출 소자 등과 같은 분야에 매우 높은 응용 가능성을 가질 것으로 기대된다.

• 생약학회지
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• 제17권2호
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• pp.134-138
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• 1986

Based on a tracer study, starch phosphorylase was implicated as an agent in the starch synthesis in sweet potato roots. The enzyme was purified from the tissue as a cluster of isozymes with an average mw of 205K (fresh roots) or 159K (roots stored for 3 mon.). On SDS polyacrylamide gel electrophoresis, one large subunit of 98K mw and several small ones of 47${\sim}57K mw were observed. From the mw data and the results of peptide mapping and immunoelectrophoretic blotting using mono- and polyclonal antibodies, it was deduced that a large part of the large subunit was cleaved at the middle part of the peptide chain to give rise to the small subunits, and on storage, the enzyme molecules were further modified by proteolysis. During the course of phosphorylase purification, a proteinaceous inhibitor of the enzyme was isolated. It had a mw of 250K and was composed of 5 identical subunits of 51K mw. In the direction of starch synthesis, the inhibitor showed a noncompetitive kinetics with a Ki of $1.3{\times}10^{-6}\;M$. By immunohistochemical methods, both the enzyme and the inhibitor were located on the cell wall and amyloplast. Crossreacting materials of the inhibitor were present in spinach leaf, potato tuber and rice grain. These findings indicate the wide occurrence of the inhibitor and also imply its possible participation in regulating starch phosphorylase activity in vivo.

• Bulletin of the Korean Chemical Society
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• 제32권8호
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• pp.2549-2552
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• 2011

In this study, the kinetic properties of wild-type and C117D mutant H. influenzae MurA (Hi MurA), which catalyzes the first reaction in the biosynthetic pathway of the cell wall, were characterized. Purified recombinant Hi MurA was active at pH values ranging from pH 5.5 to pH 10, and its $K_m$ (UNAG), $K_m$ (PEP), and $k_{cat}$ values were measured to be 31 ${\mu}M$, 24 ${\mu}M$, and 210 $min^{-1}$, respectively. Hi MurA activity was effectively inhibited by fosfomycin with an $IC_{50}$ value of 60 ${\mu}M$. Hi MurA contains a cysteine residue (C117) at the loop region near the PEP binding, whereas MurA from fosfomycin resistant Mycobaterium tuberculosis or Chlamydia trachomatis contain an aspartate residue instead of the cysteine at the corresponding site. Aspartate substitution of Cys117 in Hi MurA shifted its optimum pH from 7.8 to 6.0. In addition, the $K_m$ values for UNAG and PEP were increased to 160 ${\mu}M$ and 150 ${\mu}M$, respectively, and the $k_{cat}$ value was significantly reduced to 41 $min^{-1}$. Furthermore, the C117D mutant form of Hi MurA was not inhibited by 1 mM fosfomycin. These results indicate that the Cys117 of Hi MurA is the binding site of fosfomycin and plays an important role in the fast turnover of the catalytic reaction.

• 한국미생물·생명공학회지
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• 제24권5호
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• pp.529-533
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• 1996

Melittin (ME) from honeybee venom has a broad range of strong antimicrobial activity, but it has hemolytic activity against eukaryotic cells. In order to design peptides with powerful antifungal activity without cytotoxic property of ME and understand structure-antifungal activity relationships, the hybrid peptides derived from the sequences of ME and cecropin A (CA) or magainin 2 (MA), MA(10-17)ME(1-12) and CA(1-8)ME(1-12). were synthesized by solid phase method. MA(10-17)ME(1-12) showed potent antifungal activity comparable to ME against Fusarium oxysporum with no hemolytic activity against human red blood cells. The hybrid peptides showed strong inhibi- tion of (1, 3)-$\beta$-D-glucan synthase. This result indicates that the antifungal activity of the hybrid peptides against Fusarium oxysporum is attributed to the inhibition of cell wall synthesis. The results therefore showed a successful design of a peptide having antifungal activity without hemolytic property.

• Journal of Microbiology and Biotechnology
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• 제29권5호
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• pp.758-764
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• 2019

${\beta}$-Glucan is a chief structural polymer in the cell wall of yeast. ${\beta}$-Glucan has attracted intensive attention because of its wide applications in health protection and cosmetic areas. In the present study, the ${\beta}$-glucan biosynthesis pathway in S. Cerevisiae was engineered to enhance ${\beta}$-glucan accumulation. A newly identified bacterial ${\beta}-1$, 6-glucan synthase GsmA from Mycoplasma agalactiae was expressed, and increased ${\beta}$-glucan content by 43%. In addition, other pathway enzymes were investigated to direct more metabolic flux towards the building of ${\beta}$-glucan chains. We found that overexpression of Pgm2 (phosphoglucomutase) and Rho1 (a GTPase for activating glucan synthesis) significantly increased ${\beta}$-glucan accumulation. After further optimization of culture conditions, the ${\beta}$-glucan content was increased by 53.1%. This study provides a new approach to enhance ${\beta}$-glucan biosynthesis in Saccharomyces cerevisiae.

• 식물조직배양학회지
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• 제21권5호
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• pp.253-275
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• 1994

Many flowering plants possess genetically controlled self -incompatibility (SI) system that prevents inbreeding and promotes outcrosses. SI is usually controlled by a single, multiallelic S-locus. In gametophytically controlled system, SI results when the S-allele of the pollen is matched by one of the two S-alleles in the style, while in the sporophytic system self-incompatible reaction occurs by the interaction between the pistil genotype and genotype of, not the pollen, but the pollen parent In the former system the self-incompatible phenotype of pollen is determined by the haploid genome of the pollen itself but in the latter the pollen phenotype is governed by the genotype of the pollen parent along with the occurrence of either to-dominant or dominant/recessive allelic interactions. In the sporophytic type the inhibition reaction occurs within minutes following pollen-stigma contact, the incompatible pollen grains usually failing to germinate, whereas in gametophytic system pollen tube inhibition takes place during growth in the transmitting tissue of the style. Recognition and rejection of self pollen are the result of interaction between the S-locus protein in the pistil and the pollen protein. In the gametophytic SI the S-associated glycoprotein which is similar to the fungal ribonuclease in structure and function are localized at the intercellular matrix in the transmitting tissue of the style, with the highest concentration in the collar of the stigma, while in the sporophytic SI deposit of abundant S-locus specific glycoprotein (SLSG).is detected in the cell wall of stigmatic papillae of the open flowers. In the gametophytic system S-gene is expressed mostly at the stigmatic collar the upper third of the style length and in the pollen after meiosis. On the other hand, in the sporophytic SI S-glycoprotein gene is expressed in the papillar cells of the stigma as well as in e sporophytic tape is cells of anther wall. Recognition and rejection of self pollen in the gametophytic type is the reaction between the ribonuclease in the transmitting tissue of the style and the protein in the cytoplasm of pollen tube, whereas in the sporophytic system the inhibition of selfed pollen is caused by the interaction between the Sycoprotein in the wall of stigmatic papillar cell and the tapetum-origin protein deposited on the outer wall of the pollen grain. The claim that the S-allele-associated proteins are involved in recognition and rejection of self pollen has been made merely based on indirect evidence. Recently it has been verified that inhibition of synthesis of S$_3$ protein in Petunia inflata plants of S$_2$S$_3$ genotype by the antisense S$_3$ gene resulted in failure of the transgenic plant to reject S$_3$ pollen and that expression of the transgenic encoding S$_3$ protein in the S$_1$S$_2$ genotype confers on the transgenic plant the ability to reject S$_3$ pollen. These finding Provide direct evidence that S-proteins control the s elf-incompatibility behavior of the pistil.

• Journal of Microbiology and Biotechnology
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• 제28권3호
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• pp.482-490
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• 2018

Candida albicans infections are often problematic to treat owing to antifungal resistance, as such infections are mostly associated with biofilms. The ability of C. albicans to switch from a budding yeast to filamentous hyphae and to adhere to host cells or various surfaces supports biofilm formation. Previously, the ethanol extract from Paeonia lactiflora was reported to inhibit cell wall synthesis and cause depolarization and permeabilization of the cell membrane in C. albicans. In this study, the P. lactiflora extract was found to significantly reduce the initial stage of C. albicans biofilms from 12 clinical isolates by 38.4%. Thus, to assess the action mechanism, the effect of the P. lactiflora extract on the adhesion of C. albicans cells to polystyrene and germ tube formation was investigated using a microscopic analysis. The density of the adherent cells was diminished following incubation with the P. lactiflora extract in an acidic medium. Additionally, the P. lactiflora-treated C. albicans cells were mostly composed of less virulent pseudohyphae, and ruptured debris was found in the serum-containing medium. A quantitative real-time PCR analysis indicated that P. lactiflora downregulated the expression of C. albicans hypha-specific genes: ALS3 by 65% (p = 0.004), ECE1 by 34.9% (p = 0.001), HWP1 by 29.2% (p = 0.002), and SAP1 by 37.5% (p = 0.001), matching the microscopic analysis of the P. lactiflora action on biofilm formation. Therefore, the current findings demonstrate that the P. lactiflora ethanol extract is effective in inhibiting C. albicans biofilms in vitro, suggesting its therapeutic potential for the treatment of biofilm-associated infections.

• 한국수산과학회지
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• 제31권4호
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• pp.560-566
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• 1998

Lactobacillus sp. GM7311이 생산하는 박테리오신의 Gram 양성균에 대한 작용 형태를 알아보기 위하여, 세가지 시험균주의 TSB 배양액에 일정한 시간 간격별로 $100\;Bu/m{\ell}$ 되도록 박테리오신을 첨가한 후 그 균수 변화와 작용 특성을 살펴보았다. 우선, 시험 균주의 증식 시기별로 박테리오신을 첨가한 경우 Listeria monocytogenes는 대수증식기 후기 이후에 억제작용을 나타낸 반면, Bacillus subtilis의 경우는 대수증식기 중기 이전까지의 시험균에 대해서 강한 억제작용을 보였으며, Staphylococcus aureus는 정지기 이전의 시험균에 대해 큰 항균 효과를 나타냈다 또한 박테리오신을 처리한 세 가지 시험 균주를 전자 현미경으로 관찰했을 때, L. monocpogenes와 B. subtilis는 세포벽 파괴에 의한 세포내 물질의 유출이 관찰되었으나, S. aureus는 다른 두 균주와 달리 박테리오신에 의한 세포벽의 파괴 현상은 없었으며 다만 세포벽이 대조구에 비해 엷어진 것이 관찰되었다. 아미노산 및 지방산 조성의 변화를 조사하여 박테리오신을 처리하지 않은 대조구와 비교하였을 때, 아미노산의 경우는 세 균주 모두에서 대부분의 성분이 감소하는 경향을 보였고 지방산은 큰 조성상의 변화나 공통된 특정지방산 성분의 감소 등은 나타나지 않았으며 각 성분들의 함량이 증감하는 경향을 보였다.

• Molecules and Cells
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• 제45권9호
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• pp.660-672
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• 2022

The target of rapamycin complex (TORC) plays a key role in plant cell growth and survival by regulating the gene expression and metabolism according to environmental information. TORC activates transcription, mRNA translation, and anabolic processes under favorable conditions, thereby promoting plant growth and development. Tomato fruit ripening is a complex developmental process promoted by ethylene and specific transcription factors. TORC is known to modulate leaf senescence in tomato. In this study, we investigated the function of TORC in tomato fruit ripening using virus-induced gene silencing (VIGS) of the TORC genes, TOR, lethal with SEC13 protein 8 (LST8), and regulatory-associated protein of TOR (RAPTOR). Quantitative reverse transcription-polymerase chain reaction showed that the expression levels of tomato TORC genes were the highest in the orange stage during fruit development in Micro-Tom tomato. VIGS of these TORC genes using stage 2 tomato accelerated fruit ripening with premature orange/red coloring and decreased fruit growth, when control tobacco rattle virus 2 (TRV2)-myc fruits reached the mature green stage. TORC-deficient fruits showed early accumulation of carotenoid lycopene and reduced cellulose deposition in pericarp cell walls. The early ripening fruits had higher levels of transcripts related to fruit ripening transcription factors, ethylene biosynthesis, carotenoid synthesis, and cell wall modification. Finally, the early ripening phenotype in Micro-Tom tomato was reproduced in the commercial cultivar Moneymaker tomato by VIGS of the TORC genes. Collectively, these results demonstrate that TORC plays an important role in tomato fruit ripening by modulating the transcription of various ripening-related genes.