• The Plant Pathology Journal
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• 제35권3호
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• pp.208-218
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• 2019

Here, we reported a novel secreted protein elicitor PeBL2 from Brevibacillus laterosporus A60, which can induce hypersensitive response in tobacco (Nicotiana benthamiana). The ion-exchange chromatography, high-performance liquid chromatography (HPLC) and mass spectrometry were performed for identification of protein elicitor. The 471 bp PeBL2 gene produces a 17.22 kDa protein with 156 amino acids containing an 84-residue signal peptide. Consistent with endogenous protein, the recombinant protein expressed in Escherichia coli induced the typical hypersensitive response (HR) and necrosis in tobacco leaves. Additionally, PeBL2 also triggered early defensive response of generation of reactive oxygen species ($H_2O_2$ and $O_2{^-}$) and systemic resistance against of B. cinerea. Our findings shed new light on a novel strategy for biocontrol using B. laterosporus A60.

• BMB Reports
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• 제39권3호
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• pp.311-318
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• 2006

One of the biggest group of proteins influenced by protein kinase CK2 is formed by factors engaged in gene expression. Here we have reported recently identified yeast transcription elongation factor Elf1 as a new substrate for both monomeric and tetrameric forms of CK2. Elf1 serves as a substrate for both the recombinant CK2$\alpha$' ($K_m$ 0.38 ${\mu}M$) and holoenzyme ($K_m$ $0.13\;{\mu}M$). By MALDI-MS we identified the two serine residues at positions 95 and 117 as the most probable in vitro phosphorylation sites. Co-immunoprecypitation experiments show that Elf1 interacts with catalytic ($\alpha$ and $\alpha$') as well as regulatory ($\beta$ and $\beta$') subunits of CK2. These data may help to elucidate the role of protein kinase CK2 and Elf1 in the regulation of transcription elongation.

• The Plant Pathology Journal
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• 제35권2호
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• pp.91-99
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• 2019

Mitogen-activated protein kinase (MAPK) cascades in fungi are ubiquitously conserved signaling pathways that regulate stress responses, vegetative growth, pathogenicity, and many other developmental processes. Previously, we reported that the AbSte7 gene, which encodes a mitogen-activated protein kinase kinase (MAPKK) in Alternaria brassicicola, plays a central role in pathogenicity against host cabbage plants. In this research, we further characterized the role of AbSte7 in the pathogenicity of this fungus using ${\Delta}AbSte7$ mutants. Disruption of the AbSte7 gene of A. brassicicola reduced accumulation of metabolites toxic to the host plant in liquid culture media. The ${\Delta}AbSte7$ mutants could not efficiently detoxify cruciferous phytoalexin brassinin, possibly due to reduced expression of the brassinin hydrolase gene involved in detoxifying brassinin. Disruption of the AbSte7 gene also severely impaired fungal detoxification of reactive oxygen species. AbSte7 gene disruption reduced the enzymatic activity of cell walldegrading enzymes, including cellulase, ${\beta}$-glucosidase, pectin methylesterase, polymethyl-galacturonase, and polygalacturonic acid transeliminase, during host plant infection. Altogether, the data strongly suggest the MAPKK gene AbSte7 plays a pivotal role in A. brassicicola during host infection by regulating multiple steps, and thus increasing pathogenicity and inhibiting host defenses.

• Asian-Australasian Journal of Animal Sciences
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• 제19권8호
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• pp.1164-1173
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• 2006

Cellulase from Aspergillus niger, (${\alpha}$-amylase from Bacillus sp. and protease from Bacillus globigii were used as enzyme sources in this study to examine how their respective substrates protect them in two kinds of simulated gastrointestinal tract digesting processes. Avian total digest tract simulation test showed that filter paper, Avicel and cellulose resulted in 7.7, 6.4 and 7.4 times more activity than of unprotected cellulose, respectively. Protease with addition of casein, gelatin or soybean protein showed no significant protection response. Starch protected amylase to be 2.5 times activity of the unprotected one. Monogastric animal total tract digestion simulation test showed that filter paper, Avicel and cellulose resulted in 5.9, 9.0 and 8.8 times activity of unprotected cellulase, respectively. Casein, gelatin and soybean protein resulted in 1.2, 1.3 and 2.0 times activity of unprotected protease, respectively. Starch did not protect amylase activity in monogastric animal total tract simulation. Protection of mixed enzymes by substrates in two animal total tract simulation tests showed that filter paper in combination with soybean protein resulted in 1.5 times activity of unprotected cellulose, but all substrates tested showed no significant protection effect to protease. Soybean protein and starch added at the same time protected the amylase activity to be two times of the unprotected one. Test of non-purified substrate protection in two animal total digest tract simulation showed that cellulase activity increased as BSA (bovine serum albumin) concentration increased, with the highest activity to be 1.3 times of unprotected enzyme. However, BSA showed no significant protection effect to protease. Amylase activity increased to 1.5 times as BSA added more than 1.5% (w/v). Cellulase activity increased to 1.5 times as soybean hull was added higher than 1.5%. Amylase had a significant protection response only when soybean hull added up to 2%. Protease activity was not protected by soybean hull to any significant extent.

• Asian-Australasian Journal of Animal Sciences
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• 제13권4호
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• pp.535-542
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• 2000

Canola meal and seed are poor sources of ruminal undegraded protein (RUP). On average, canola meal and canola seed contains 35 and 14% RUP, respectively. Several protection methods are effective in reducing ruminal degradation of canola protein and in increasing RUP without affecting total tract protein digestibility. Heat (e.g., dry heat, moist heat and jet-sploding) and chemical (e.g., formaldehyde) treatments are the most common methods used to reduce ruminal degradability of canola protein. In most cases, heat treatments were found to be more effective than chemical treatments in protecting canola protein form ruminal degradation. Despite improvement in RUP content and intestinal availability of RUP, data form several studies showed little or no improvement in animal performance as a result of increasing the RUDP level of canola meal and seed.

• 생명과학회지
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• 제29권9호
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• pp.949-954
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• 2019

세포막 단백질의 topology는 막단백질의 중요한 특징이다. 우리는 이전에 C4orf32단백질을 클로닝 하였으나, 이 단백질의 세포내 위치나 topology는 알지 못했다. 이번 연구를 통해 C4orf32는 세포내에서 소포체에 위치되는 막단백질임을 알게 되었다. C4orf32의 topology를 알기 위해 protease protection assay, fluorescence protease protection (FPP) assay, FRB/rapamycin/FKBP system을 활용하였다. Protease protection assay와 FPP assay를 적용한 결과 C-말단에 GFP를 붙인 C4orf32-GFP의 경우 GFP가 소포체의 세포질 표면에 위치함을 확인할 수 있었다. 또한, FRB/rapamycin/FKBP시스템을 이용한 실험에서 rapamycin이 처리되지 않은 경우는 mRFP-FKBP가 세포질에 위치하다가 rapamycin이 처리되면 C4orf32-GFP-FR가 위치하는 소포체로 이동함을 확인할 수 있었다. 이러한 사실은 C4orf32의 C-말단이 소포체의 세포질쪽 면에 위치한다는 사실을 말해준다. 이러한 연구를 통해 C4orf32는 Type I 소포체 막단백질에 속한다는 사실을 확인할 수 있었다.

• The Plant Pathology Journal
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• 제36권4호
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• pp.323-334
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• 2020

Lysin motif (LysM) proteins are reported to be necessary for the virulence and immune response suppression in many herbaceous plant pathogens, while far less is documented in woody plant pathogens. In this study, we preliminarily characterized the molecular function of a LysM protein LtLysM1 in woody plant pathogen Lasiodiplodia theobromae. Transcriptional profiles revealed that LtLysM1 is highly expressed at infectious stages, especially at 36 and 48 hours post inoculation. Amino acid sequence analyses revealed that LtLysM1 was a putative glycoprotein with 10 predicted N-glycosylation sites and one LysM domain. Pathogenicity tests showed that overexpressed transformants of LtLysM1 displayed increased virulence on grapevine shoots in comparison with that of wild type CSS-01s, and RNAi transformants of LtLysM1 exhibited significantly decreased lesion length when compared with that of wild type CSS-01s. Moreover, LtLysM1 was confirmed to be a secreted protein by a yeast signal peptide trap assay. Transient expression in Nicotiana benthamiana together with protein immunoblotting confirmed that LtLysM1 was an N-glycosylated protein. In contrast to previously reported LysM protein Slp1 and OsCEBiP, LtLysM1 molecule did not interact with itself based on yeast two hybrid and co-immunoprecipitation assays. These results indicate that LtLysM1 is a secreted protein and functions as a critical virulence factor during the disease symptom development in woody plants.

• Korea Journal of Cosmetic Science
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• 제2권1호
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• pp.1-10
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• 2020

Protection mechanisms for skin damage of ultraviolet (UV) absorbers in personal care products for protection against UV are well studied, but not for hair protection. The purpose of this study is to describe and compare the changes of physical property produced in human hair by doses of the UV-B exposure causing protein degradation. To observe the change of physical properties in hair, the experimental intensity of UV-B exposure has been established on the basis of statistical data from official meterological administration as daily one hour sunlight exposure for two weeks. Polysilicone-15, ethylhexyl methoxycinnamate (OMC), and octocrylene were employed for UV-B absorber, and those were treated to hair swatch by rubbing wash through shampoo and conditioner. Bending rigidity displayed kinetically successive reduction at high doses of UV exposure up to the 8,000 s, and exhibited different level at each sample of UV-B absorber. However, the values of Bossa Nova Technologies (BNT) for shinning factor were already saturable at the 2,000 s exposure except that treated with polysilicone-15. The differential scanning calorimetry (DSC) to measure a strength of inner protein produces a successive reduction of enthalpy as like a reduction of bending rigidity upon UV exposure. Surface roughness from lateral force microscope (LFM) acquired immediately after UV exposure show a saturable frictional voltage which has been also found in a saturable BNT data as the time of UV exposure increases. Through researching the DSC and the LFM, shinning of hair was much correlated to the protein damage at the surface, and bending rigidity could be regulated by the protein structural damage inside hair. Therefore, the optimization of efficient strategy for simultaneous prevention of hair protein on the surface and internal hair was required to maintain physical properties against UV.

• Journal of Radiation Protection and Research
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• 제20권2호
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• pp.71-78
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• 1995

본 연구에서는 유전자 손상회복에 관여하는 단백질을 이용하여 돌연변이 생성을 억제시키는 물질로서 알려진 cobaltous chloride가 유전자 손상회복에 미치는 영향을 연구하므로서 방사선으로 인한 손상방지 및 방사선 방어효과에 대한 적용가능성을 평가하였다. Cobaltous chloride가 RecA 단백질의 기능에 미치는 영향을 조사한 결과 RecA 단백질에 의한 DNA strand exchange 반웅에 있어 cobaltous chloride 처리로 RecA 단백질이 $_{ss}DNA$로 부터 SSB 단백질과 더 효과적으로 경쟁함으로써 안정된 $RecA-_{ss}DNA$ complex의 형성을 유도하고, 증가된 ATPase활성에 의한 ATP 가수분해로 손상된 DNA의 회복이 촉진될 수 있다는 사실을 입증 해주고 있다. 또한 RecA단백질은 UV에 의해 손상된 supercoiled DNA에 더 효과적으로 결합됨이 관찰되었으며 UV 선량과도 상관관계가 있음을 확인하였다. 따라서 이와 같은 연구결과들은 방사선으로 인한 유전적인 손상방지 및 방사선 방어효과에 관한 연구에 적용될 수 있을 것으로 기대된다.

• The Plant Pathology Journal
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• 제35권5호
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• pp.498-507
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• 2019

Odontoglossum ringspot virus (ORSV) is a member of the genus Tobamovirus. It is one of the most prevalent viruses infecting orchids worldwide. Earlier studies reported the genetic variability of ORSV isolates from Korea and China. However, the evolutionary rate, timescale, and phylogeographical analyses of ORSV were unclear. Twenty-one coat protein (CP) gene sequences of ORSV were determined in this study, and used them together with 145 CP sequences obtained from GenBank to infer the genetic diversities, evolutionary rate, timescale and migration of ORSV populations. Evolutionary rate of ORSV populations was $1.25{\times}10^{-3}nucleotides/site/y$. The most recent common ancestors came from 30 year ago (95% confidence intervals, 26-40). Based on CP gene, ORSV migrated from mainland China and South Korea to Taiwan island, Germany, Australia, Singapore, and Indonesia, and it also circulated within east Asia. Our study is the first attempt to evaluate the evolutionary rates, timescales and migration dynamics of ORSV.