• The Plant Pathology Journal
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• 제35권5호
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• pp.530-537
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• 2019

Fuji, a major apple cultivar in Korea, is susceptible to white rot. Apple white rot disease appears on the stem and fruit; the development of which deteriorates fruit quality, resulting in decreases in farmers' income. Thus, it is necessary to characterize molecular markers related to apple white rot resistance. In this study, we screened for differentially expressed genes between uninfected apple fruits and those infected with Botryosphaeria dothidea, the fungal pathogen that causes white rot. Antimicrobial tests suggest that a gene expression involved in the synthesis of the substance inhibiting the growth of B. dothidea in apples was induced by pathogen infection. We identified seven transcripts induced by the infection. The seven transcripts were homologous to genes encoding a flavonoid glucosyltransferase, a metallothionein-like protein, a senescence-induced protein, a chitinase, a wound-induced protein, and proteins of unknown function. These genes have functions related to responses to environmental stresses, including pathogen infections. Our results can be useful for the development of molecular markers for early detection of the disease or for use in breeding white rotresistant cultivars.

• 한국자원식물학회지
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• 제27권4호
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• pp.392-399
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• 2014

본 연구에서는 식물의 노쇠와 관련된 SENESCENCE 1 (SEN1) 유전자를 현사시나무에서 분리하고, 여러 가지 조건에서 발현 특성을 분석하였다. 현사시나무의 SEN1 유전자(PagSEN1)는 243개의 아미노산으로 이루어져 있고, 한 개의 rhodanese domain을 가지고 있다. Southern blot 분석 결과 PagSEN1 유전자는 현사시나무에서 2 copy 정도가 존재하는 것으로 나타났다. 조직 특이적 발현양상을 분석한 결과 PagSEN1 유전자는 성숙잎에서 가장 높게 발현하고, 뿌리에서 가장 낮게 발현하는 것으로 나타났다. 또한 mannitol과 염 스트레스 처리에 의해 300배 이상 발현이 증가한 반면에 저온 스트레스에는 반응하지 않았다. 식물 호르몬 처리에서는 ABA와 JA 처리 10시간 후에 발현이 3.5배와 2.4배 이상 증가하는 것으로 나타났다. 따라서 PagSEN1 유전자는 건조와 염 스트레스에 반응하며, 식물의 자연적 노쇠과정 뿐 아니라 스트레스와 같은 환경변화에 의해 유발되는 노쇠과정에도 관여하는 것으로 판단된다.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.78.1-78
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• 2003

An osmotin-like protein (CAOSMl) gene was isolated from pepper leaves infected with the avirulent strain Bv5-4a of Xmthomonas campestris pv. vesicatoria. The cDNA encodes a polypeptide of 250 amino acids with a molecular mass of 27, 361 Da. Its amino acid sequence is highly homologous to various osmotin-like proteins from other plant species. The CAOSMl gene expression was organ- and tissue-specifically regulated In pepper plants. The CAOSMl mRNA was intensely localized in the endodermis area of root tissue and in the phloem cells of vascular bundles of red fruit tissue, but not in leaf, stem, and green fruit tissues of healthy pepper plants. Infection by X. c. pv vesintoria, Colletotrichum coccodes, or Phytopkhora capsici iinduced CAOSMl transcription in the leaf or stem tissues. Expression of the CAOSMl gene was somewhat higher in the incompatible than the compatible interactions of pathogens with pepper. The CAOSMl mRNA was prevalently localized in the phloem cells of the vascular bundle of leaf tissues infected by C. coccodes. The CAOSMl gene was activated in leaf tissues by treatment with ethylene, methyl jasmonate, high salinity, cold acclimation and mechanical wounding, but not by abscisic acid (ABA) and drought. These results indicate that the pepper CAOSMl protein functions in response to Pathogens and some abiotic stresses in pepper plants

• Journal of Plant Biotechnology
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• 제36권4호
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• pp.407-414
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• 2009

A cDNA clone encoding phytocystatin was isolated from Brassica rapa seedlings, through rapid amplification of cDNA ends (RACE). This gene (name as Brcpi1; GenBank accession no.: EF079953) had a total length of 881 bp with an open reading frame of 609 bp, and encoded predicted polypeptide of 203 amino acid (aa) residues including a putative N-terminal signal peptide. Other relevant regions found its sequence included the G and PW conserved aa motifs, and the consensus LARFAV sequence for phytocystatins and the reactive site QVVAG. The BrCPI1 protein shared 95, 94, 81, 80 and 78% identity with other CPI proterins isolated from Brassica oleracea (BoCPI-1), Arabidopsis thaliana (AtCY SB), Glycine max (GmCPI), Oryza sativa (OsCYS-2) and Zea may (ZmCPI) at amino acid level, respectively. Southern blot analysis showed that Brcpi1 was a low copy gene. Expression pattern analysis revealed that Brcpi1 was a tissue-specific expressing gene during reproductive growth and strongly expressed at mature seedling stages. Furthermore, overexpression of Brcpi1 in transgenic Arabidopsis was enhanced tolerance to salt and cold stresses. Meanwhile the juvenile seedling of Brcpi1 transgenic plants was not affected by various concentrations ABA in MS medium. Taken together, the results showed that Brcpi1 functioned as a cysteine protease inhibitor and it exhibited a protective agent against diverse types of abiotic stress, which induced this gene in a tissue- and stress-specific manner.

• BMB Reports
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• 제38권4호
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• pp.440-446
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• 2005

A cDNA that was rapidly induced upon abscisic acid, cold, drought, mechanical wounding and to a lesser extent, by high salinity treatment, was isolated from Arabidopsis seedlings. It was classified as DREB subfamily member based on multiple sequence alignment and phylogenetic characterization. Since it encoded a protein with a typical ERF/AP2 DNA-binding domain and was closely related to the TINY gene, we named it TINY2. Gel retardation assay revealed that TINY2 was able to form a specific complex with the previously characterized DRE element while showed only residual affinity to the GCC box. When fused to the GAL4 DNA-binding domain, either full-length or its C-terminus functioned effectively as a trans-activator in the yeast one-hybrid assay while its N-terminus was completely inactive. Our data indicate that TINY2 could be a new member of the AP2/EREBP transcription factor family involved in activation of down-stream genes in response to environmental stress.

• Molecules and Cells
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• 제41권10호
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• pp.900-908
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• 2018

Insulin resistance is closely associated with metabolic diseases such as type 2 diabetes, dyslipidemia, hypertension and atherosclerosis. Thiazolidinediones (TZDs) have been developed to ameliorate insulin resistance by activation of peroxisome proliferator-activated receptor (PPAR) ${\gamma}$. Although TZDs are synthetic ligands for $PPAR{\gamma}$, metabolic outcomes of each TZD are different. Moreover, there are lack of head-to-head comparative studies among TZDs in the aspect of metabolic outcomes. In this study, we analyzed the effects of three TZDs, including lobeglitazone (Lobe), rosiglitazone (Rosi), and pioglitazone (Pio) on metabolic and thermogenic regulation. In adipocytes, Lobe more potently stimulated adipogenesis and insulin-dependent glucose uptake than Rosi and Pio. In the presence of pro-inflammatory stimuli, Lobe efficiently suppressed expressions of pro-inflammatory genes in macrophages and adipocytes. In obese and diabetic db/db mice, Lobe effectively promoted insulin-stimulated glucose uptake and suppressed pro-inflammatory responses in epididymal white adipose tissue (EAT), leading to improve glucose intolerance. Compared to other two TZDs, Lobe enhanced beige adipocyte formation and thermogenic gene expression in inguinal white adipose tissue (IAT) of lean mice, which would be attributable to cold-induced thermogenesis. Collectively, these comparison data suggest that Lobe could relieve insulin resistance and enhance thermogenesis at low-concentration conditions where Rosi and Pio are less effective.

• 한국축산식품학회지
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• 제33권1호
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• pp.75-82
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• 2013

This study aimed to isolate lactic acid bacteria (LAB) from Kimchi and to identify suitable probiotic strain for application in fermented dairy product as a commercial starter culture. A total of 106 (LAB) strains were isolated from Kimchi collected from different regions in Korea and their phenotypic characteristics were assayed. Four isolates from MRS agar plates were selected and designated as DKL109, DKL119, DKL121 and DKL128. They were identified first by API 50 CHL kit and then 16S rRNA gene sequencing. DKL121 and DKL128 were identified as Lactobacillus paracasei and Lactobacillus casei, respectively. Other two isolates (DKL109 and DKL119) were identified as Lactobacillus plantarum. To estimate their applicability in dairy products, the characteristics including acid and bile tolerance, cold shock induced cryotolerance and enzymatic activities were determined. There was wide variation in ability of strains to acid tolerance, but no significant differences in bile tolerance, cold shock induced cryotolerance within selected strains. DKL119 and DKL121 showed the highest resistance to acid and bile and the highest ${\beta}$-galactosidase activity, respectively. When these two strains were used for yogurt preparation as a single starter culture, their viable cell counts reached to $1.0{\times}10^9CFU/mL$. Lactobacillus plantarum DKL119 showed faster acid development than commercial starter culture. Also storage trials at $10^{\circ}C$ showed that the viability of these strains was retained over 15 d. With these results, it was indicated that probiotics isolated from Kimchi can be used in yogurt manufacturing as a starter culture.

• 한국미생물·생명공학회지
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• 제51권2호
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• pp.191-202
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• 2023

Elevated serum cholesterol is a main risk factor for heart disorders. Most probiotic products administered to lower cholesterol are dairy products which are not suitable for lactose-intolerant individuals. In this study, we assessed the cholesterol-lowering efficacy of LAB isolated from traditionally fermented drinks in diet-induced rats and determine their efficacy in the production of non-dairy, probiotic formulations using papaya juice. LAB were isolated from palm wine and corn beer on MRS agar using a pour-plate technique. Identification was carried out using 16S rRNA gene sequencing. A hypercholesterolemia model in which diet-induced Wistar albino rats were assigned into four groups was established. Oral gavage was carried out for 30 days. On the 31^st day, the rats were dissected and the serum lipid profile was analyzed using biochemical kits. A 10⁶ cfu/ml of a 24-h-old culture of selected lactobacilli was used to inoculate papaya juice and incubated at 37℃. Microbial and chemical changes were assessed during papaya fermentation and after four weeks of cold storage. Two selected isolates (Pw1 and Cb4) had in vitro cholesterol reduction of > 80%. These two isolates lowered lipid profile (triglyceride, total cholesterol, LDL-c) significantly, and increased HDL-c levels (p < 0.5) in the rat sera. Phylogenetic analysis showed that Pw1 was 98.86% similar to Limosilactobacillus fermentum, while Cb4 was 99.54% similar to Enteroccocus faecium. Both strains fermented papaya juice with cell viability reaching 8.92 × 10⁸ cfu/ml and 25.3 × 10⁸ cfu/ml respectively, and were still viable after 4 weeks of cold storage.

• Journal of Plant Biotechnology
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• 제40권2호
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• pp.102-110
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• 2013

The novel BrRZFPs genes encoding C3HC4-type RING zinc finger protein were identified from FOX (full length cDNA over-expressing) library of Brassica rapa. Ten full-length cDNAs obtained from the library encode zinc-finger protein containing 346 amino acids, designated BrRZFPs. These genes were classified into four groups by phylogenic analysis showing conserved protein sequences at both termini. The tissue distribution of BrRZFPs transcription was examined by qRT-PCR revealing ubiquitous expression pattern. However, each gene was strongly expressed in the specific tissue. Transcriptional analysis showed that those acquired 10 genes were inducible under abiotic stresses. Likewise, the transcript of BrRZFP3 was strongly induced (~12-folds) by exogenous abscisic acid, whereas the transcripts of BrRZFP1, BrRZFP2 and BrRZFP3 were (> 9-folds) induced by cold. We suggest that these BrRZFPs that function as signal or response to abiotic stress are useful for crop improvement.

• 생명과학회지
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• 제25권5호
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• pp.507-514
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• 2015

D-Xylulose는 nonoxidative pentose phosphate 경로를 통해 glycolysis 과정으로 들어가기 전에 D-xylulose kinase에 의해서 D-xylulose-5-phosphate로 인산화 된다. K. gwangalliensis strain SJ2로부터 D-xylulose kinase (XK)를 암호화하는 유전자는 E. coli를 이용하여 서열분석 및 발현 하였으며, XK 유전자의 염기서열 1,419 bp로 구성되어 있으며 463개의 아미노산 잔기를 암호화하고 있다. 분석결과를 통해 XK 유전자가 진화과정 동안 잘 보존되었음을 보여 주었다. XK 유전자의 발현을 위해 pCold-II 발현 벡터에 클로닝 하였으며 클로닝 된 플라스미드는 E. coli strain BL21 (DE3)에 형질전환 하여 IPTG를 이용해 발현을 유도하였다. 재조합 된 XK 단백질의 크기는 약 48 kDa이었다. 이 발현된 단백질은 affinity chromatography를 이용하여 정제하였으며 D-xylulose kinase에 따른 enzymatic activity를 분석하였다. D-xylulose와 ATP로 실행한 XK enzyme kinetic 연구는 각각 250±20 μM과 1,300±50 μM의 Km value를 보였다. 본 연구를 통해 얻어진 결과는 분자적 수준에서 D-xylulose kinase의 특성연구의 보다 넓은 지식적 기초를 제공할 것으로 사료된다.