• Title/Summary/Keyword: 유전자 서열 분석

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Identification and Chemotype Profiling of Fusarium Head Blight Disease in Triticale (국내 재배 트리티케일에 발생한 붉은곰팡이병의 다양성 및 독소화학형 분석)

  • Yang, Jung-Wook;Kim, Joo-Yeon;Lee, Mi-Rang;Kang, In-Jeong;Jeong, Jung-Hyun;Park, Myoung Ryoul;Ku, Ja-Hwan;Kim, Wook-Han
    • Research in Plant Disease
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    • v.27 no.4
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    • pp.172-179
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    • 2021
  • This study aimed to assess the disease incidence and distribution of toxigenic in Korean triticale. The pathogen of triticale that cause Fusarium head blight were isolated from five different triticale cultivars that cultivated in Suwon Korea at 2021 year. The 72 candidate were classified as a Fusarium asiaticum by morphology analysis and by ITS1, TEF-1α gene sequence analysis. And the results of pathogenicity with 72 isolates on seedling triticale, 71 isolates were showed disease symptom. Also, seven out of 71 Fusarium isolates were inoculated on the wheat, to test the pathogenicity on the different host. The results showed more low pathogenicity on the wheat than triticale. The results of analysis of toxin type with 72 isolates, 64.6% isolates were produced nivalenol type toxin and other 4.6% and 30.8% isolates were produce 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol, respectively. To select fungicide for control, the 72 Fusarium isolates were cultivated on the media that containing four kinds fungicide. The captan, hexaconazole, and difenoconazole·propiconazole treated Fusarium isolates were not showed resistance response against each fungicide. However, six isolates out of 72 isolates, showed resistance response to fludioxonil. This study is first report that F. asiaticum causes Fusarium head blight disease of triticale in Korea.

Characterization of typical Aeromonas salmonicida isolated from Sea-Chum Salmon (Oncorhynchus keta) (해수에 순치된 첨연어(Oncorhynchus keta)에서 분리된 정형 에로모나스 살모니시다(Aeromonas salmonicida)에 대한 특성 분석)

  • Jongwon Lim;Sungjae Ko;Youngjun Park;Do-il Ahn;Suhee Hong
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.263-275
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    • 2023
  • Chum salmon (Oncorhynchus keta) is a species which returns to Korea for spawning and was produced as seed production at the Fisheries Resources Agency located in Uljin-gun, Gyeongsangbuk-do to preserve the species. However, farmed chum salmon showed symptoms of bacterial infection. Therefore, in this study, bacteria were isolated to identify the causative agent from chum salmon in October 2021. The isolated bacteria were identified based on the sequences of 16S rDNA, rpoD (RNA polymerase sigma factor σ70), and vapA (A-layer) genes. Also, salinity-growth curve, biochemical characterization, antibiotic susceptibility test, and pathogenicity analysis were performed in four strains. As a result, four isolated strains were identified as Aeromonas salmonicida subsp. salmonicida. Additionally, the bacterial strains showed a decrease in growth as the salt concentration increased in the medium. All of the isolated strains exhibited γ-hemolysis, and the same biochemical properties. In the antimicrobial susceptibility test, all strains showed an inhibition zone of 40 to 44 mm for oxolinic acid, flumequine, and florfenicol. Pathogenic factors were assessed by RT-PCR at the mRNA level, and found that the four strains expresses the outer membrane ring of T3SS (ascV), inner membrane ring of T3SS (ascC), vapA, enterotoxin (act), and lipase (lip) genes which are well known to significantly contribute to the pathogenicity of A. salmonicida. The results of this study can be used as basic data to prevent A. salmonicida subsp. salmonicida occurring in sea-chum salmon in the future.

Optimization of Medium Components using Response Surface Methodology for Cost-effective Mannitol Production by Leuconostoc mesenteroides SRCM201425 (반응표면분석법을 이용한 Leuconostoc mesenteroides SRCM201425의 만니톨 생산배지 최적화)

  • Ha, Gwangsu;Shin, Su-Jin;Jeong, Seong-Yeop;Yang, HoYeon;Im, Sua;Heo, JuHee;Yang, Hee-Jong;Jeong, Do-Youn
    • Journal of Life Science
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    • v.29 no.8
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    • pp.861-870
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    • 2019
  • This study was undertaken to establish optimum medium compositions for cost-effective mannitol production by Leuconostoc mesenteroides SRCM201425 isolated from kimchi. L. mesenteroides SRCM21425 from kimchi was selected for efficient mannitol production based on fructose analysis and identified by its 16S rRNA gene sequence, as well as by carbohydrate fermentation pattern analysis. To enhance mannitol production by L. mesenteroides SRCM201425, the effects of carbon, nitrogen, and mineral sources on mannitol production were first determined using Plackett-Burman design (PBD). The effects of 11 variables on mannitol production were investigated of which three variables, fructose, sucrose, and peptone, were selected. In the second step, each concentration of fructose, sucrose, and peptone was optimized using a central composite design (CCD) and response surface analysis. The predicted concentrations of fructose, sucrose, and peptone were 38.68 g/l, 30 g/l, and 39.67 g/l, respectively. The mathematical response model was reliable, with a coefficient of determination of $R^2=0.9185$. Mannitol production increased 20-fold as compared with the MRS medium, corresponding to a mannitol yield 97.46% when compared to MRS supplemented with 100 g/l of fructose in flask system. Furthermore, the production in the optimized medium was cost-effective. The findings of this study can be expected to be useful in biological production for catalytic hydrogenation causing byproduct and additional production costs.

Taxonomic Characterization and Safety of Nuruk Molds Used Industrially in Korea (국내에 유통되는 종국 곰팡이의 분류학적 특성 및 안전성)

  • Hong, Seung-Beom;Hong, Sung-Yong;Jo, Kyu-Hong;Kim, Young-Sik;Do, Jong-Ho;Do, Ji-Young;Noh, Seok-Beom;Yoon, Han-Hong;Chung, Soo-Hyun
    • The Korean Journal of Mycology
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    • v.43 no.3
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    • pp.149-157
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    • 2015
  • We examined taxonomic characteristics and safety of eight Nuruk molds that are widely used for making soybean paste, soy sauce and alcoholic beverages in Korea. HK1 from Hakyeong Fermentation Co., SW101 from Suwon Fermentation Co., CF1001, CF1002, CF1003 from Chungmoo Fermaentation Co. and KACC 93210 are yellow-Nuruk molds, and SW201 from Suwon Fermentation Co. and CF1005 from Chungmoo Fermentation Co. are white-Nuruk molds. Six strains of yellow-Nuruk molds were identified as Aspergillus oryzae. HK1, SW101, CF1001 and CF1003 of yellow-Nuruk molds have middle length of stipes ($711{\sim}1,121{\mu}m$), and CF1003 (for sake) produced less conidia and more hyphae than HK1, SW101 and CF1001 (for soybean paste). CF 1002 used for soy sauce has shorter stipes ($543{\mu}m$) and is clustered into IBLB-group based on omtA gene analysis although the other yellow-Nuruk molds are clustered into ICAo group. KACC 93210 isolated from traditional Korean Meju has very short stipes (average $270{\mu}m$), and showed velvety colonies although the others showed floccose colonies. The strain has different DNA sequences of omtA gene from other strains in NCBI GenBank as well as strains used in Korea, suggesting that it is unique from other strains published. SW201 and CF1005 of white-Nuruk molds were identified as Aspergillus luchuensis or A. luchuensis mut. Kawachii that is known as safe, non-toxigenic fungus. The six strains of yellow-Nuruk molds did not produce mycotoxins including aflatoxin, cyclopiazonic acid, and sterigmatocystin. Therefore, eight strains of Nuruk molds used for making soy sauce, soybean paste and alcoholic beverages in Korea were proved to be safe in this study.

Clinical Features, Molecular Analysis, and Outcome of ERT in Korean Patients with Mucopolysaccharidosis Type VI (국내 6형 뮤코다당증의 임상 양상, 분자유전학적 특징 및 효소치료의 효과에 대한 고찰)

  • Wichajarn, Khunton;Kim, Jinsup;Yang, Aram;Sohn, Young Bae;Lee, Beom Hee;Yoo, Han-Wook;Cho, Sung Yoon;Jin, Dong-Kyu
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.16 no.1
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    • pp.24-33
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    • 2016
  • Mucopolysaccharidosis type VI (MPS VI) is a rare disease caused by the mutation of ARSB with prevalence range from 1/5,000 in northeast Brazil to 1/2,057,529 births in Czech Republic. In Asia, there is only one published figure in Taiwan of about 1/833,000 births. The exact prevalence in the Korean population is unknown, but we estimated the incidence of MPS VI is about 0.03/100,000 live births. Enzyme replacement therapy (ERT) with recombinant human Arylsulfatase B (rhASB) is a modality for the treatment of MPS VI that reduces the excretion of urine glycosaminoglycan (GAG) and improves joint motion, pulmonary function, and endurance. We presented the clinical features, molecular analysis and outcome of ERT in three Korean MPS VI patients. All patients had the typical characteristic clinical features of MPS IV. Short stature, dysostosis multiplex, corneal opacity and valvular heart disease were found at first presentation, while restrictive lung disease and carpal tunnel syndrome developed later in all patients. Molecular analysis demonstrated novel missense and nonsense mutation in the patients, including p.Ile 67Ser, p.Gly328Arg, $p.Arg191^*$, p.Asp352Asn, and p.Gly17Asp. After ERT, urine GAG was decreased in all patients. Skeletal involvement, corneal opacity, heart valve abnormalities and pulmonary function were not improved with ERT, but it had a better outcome on regarding joint motion and endurance. One patient underwent allogeneic bone marrow transplantation (BMT) prior to ERT, but their clinical response was not improved much after BMT. This study demonstrates clinical phenotypes and molecular analysis of the severe form of MPS VI in Korean patients.

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The Effect of Translationally Controlled Tumor Protein (TCTP) of the Arctic Copepod Calanus glacialis on Protecting Escherichia coli Cells against Oxidative Stress (북극 동물플랑크톤 Calanus glacialis TCTP (Translationally Controlled Tumor Protein)가 산화적 스트레스 상태에서 E. coli 세포의 저항성에 미치는 효과)

  • Park, Yu Kyung;Lee, Chang-Eun;Lee, Hyoungseok;Koh, Hye Yeon;Kim, Sojin;Lee, Sung Gu;Kim, Jung Eun;Yim, Joung Han;Hong, Ju-Mi;Kim, Ryeo-Ok;Han, Se Jong;Kim, Il-Chan
    • Journal of Life Science
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    • v.30 no.11
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    • pp.931-938
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    • 2020
  • Translationally controlled tumor protein (TCTP) is one of the most abundant proteins in various eukaryotic organisms. TCTPs play important roles in cell physiological processes in cancer, cell proliferation, gene regulation, and heat shock response. TCTP is also considered an important factor in the resistance to oxidative stress induced by dithiothreitol or hydrogen peroxide (H2O2). Arctic calanoid copepods have a variety of antioxidant defense systems to regulate the levels of potentially harmful reactive oxygen species generated by ultraviolet radiation in the Arctic marine ecosystem. However, information on the antioxidant activity of TCTP in the Arctic Calanus glacialis is still scarce. To understand the putative antioxidant function of the Arctic copepod C. glacialis TCTP (Cg-TCTP), its gene was cloned and sequenced. The Cg-TCTP comprised 522 bp and encoded a 174-amino acid putative protein with a calculated molecular weight of ~23 kDa. The recombinant Cg-TCTP (Cg-r TCTP) gene was overexpressed in Escherichia coli (BL21), and Cg-rTCTP-transformed cells were grown in the presence or absence of H2O2. Cg-rTCTP-transformed E. coli showed increased tolerance to high H2O2 concentrations. Therefore, TCTP may be an important antioxidant protein related to tolerance of the Arctic copepod C. glacialis to oxidative stress in the harsh environment of the Arctic Ocean.

Stable Inheritance of an Integrated Transgene and Its Expression in Phenylethylisothiocyanate-Enriched Transgenic Chinese cabbage (Phenylethylisothiocyanate 함량이 증진된 형질전환 배추에서의 도입유전자의 후대 유전 및 발현 안정성 검정)

  • Park, Ji-Hyun;Kim, Hyoung-Seok;Lee, Gi-Ho;Yu, Jae-Gyung;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.112-121
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    • 2016
  • Development of genetically-modified (GM) crops enables the introduction of new traits to the plant to confer characteristics such as disease resistance, herbicide resistance and human health-promoting bioactivity. Successful commercialization of newly developed GM crops requires stable inheritance of integrated T-DNA and newly introduced traits through the multiple generations. This study was carried out to confirm the stable inheritance of the integrated T-DNA in $T_1$ and $T_2$ transgenic Chinese cabbage (Brassica rapa ssp. pekinensis) that was genetically modified to increase concentrations of phenylethylisothiocyanate (PEITC), which is a potential anti-carcinogenic phytochemical. For this purpose, the IGA 1-3 ($T_1$ generation) and IGA 1-3-5 ($T_2$ generation) lines were selected by PCR and a IGA 1-3 transgenic plant ($T_1$ generation) was analyzed to confirm the T-DNA insertion site in the Chinese cabbage genome by VA-TAIL PCR. The results of this study showed that the introduced T-DNA in IGA 1 line was stably inherited to the next generations without any variations in terms of the structure of the transgenes, and this line also showed the expected transgene function that resulted in increased concentration of PEITC through the multiple generations. Finally, we confirmed the increased QR activity in IGA 1 $T_1$ and $T_2$ transgenic lines, which indicates an enhanced potential anti-carcinogenic bioactivity and its stable inheritance in IGA1 $T_1$ and $T_2$ transgenic lines.

Role of CopA to Regulate repABC Gene Expression on the Transcriptional Level (전사 수준에서 repABC 유전자 발현을 조절하는 CopA 단백질의 역할)

  • Sam Woong Kim;Sang Wan Gal;Won-Jae Chi;Woo Young Bang;Tae Wan Kim;In Gyu Baek;Kyu Ho Bang
    • Journal of Life Science
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    • v.34 no.2
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    • pp.86-93
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    • 2024
  • Since replication of plasmids must be strictly controlled, plasmids that generally perform rolling circle replication generally maintain a constant copy number by strictly controlling the replication initiator Rep at the transcriptional and translational levels. Plasmid pJB01 contains three orfs (copA, repB, repC or repABC) consisting of a single operon. From analysis of amino acid sequence, pJB01 CopA was homologous to the Cops, as a copy number control protein, of other plasmids. When compared with a CopG of pMV158, CopA seems to form the RHH (ribbon-helix-helix) known as a motif of generalized repressor of plasmids. The result of gel mobility shift assay (EMSA) revealed that the purified fusion CopA protein binds to the operator region of the repABC operon. To examine the functional role of CopA on transcriptional level, 3 point mutants were constructed in coding frame of copA such as CopA R16M, K26R and E50V. The repABC mRNA levels of CopA R16M, K26R and E50V mutants increased 1.84, 1.78 and 2.86 folds more than that of CopA wt, respectively. Furthermore, copy numbers owing to mutations in three copA genes also increased 1.86, 1.68 and 2.89 folds more than that of copA wt, respectively. These results suggest that CopA is the transcriptional repressor, and lowers the copy number of pJB01 by reducing repABC mRNA and then RepB, as a replication initiator.

Identification of Domesticated Silkworm Varieties Using a Whole Genome Single Nucleotide Polymorphisms-based Decision Tree (전장유전체 SNP 기반 decision tree를 이용한 누에 품종 판별)

  • Park, Jong Woo;Park, Jeong Sun;Jeong, Chan Young;Kwon, Hyeok Gyu;Kang, Sang Kuk;Kim, Seong-Wan;Kim, Nam-Suk;Kim, Kee Young;Kim, Iksoo
    • Journal of Life Science
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    • v.32 no.12
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    • pp.947-955
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    • 2022
  • Silkworms, which have recently shown promise as functional health foods, show functional differences between varieties; therefore, the need for variety identification is emerging. In this study, we analyzed the whole silkworm genome to identify 10 unique silkworm varieties (Baekhwang, Baekok, Daebaek, Daebak, Daehwang, Goldensilk, Hansaeng, Joohwang, Kumkang, and Kumok) using single nucleotide polymorphisms (SNP) present in the genome as biomarkers. In addition, nine SNPs were selected to discriminate between varieties by selecting SNPs specific to each variety. We subsequently created a decision tree capable of cross-verifying each variety and classifying the varieties through sequential analysis. Restriction fragment length polymorphism (RFLP) was used for SNP867 and SNP9183 to differentiate between the varieties of Daehwang and Goldensilk and between Kumkang and Daebak, respectively. A tetra-primer amplification refractory (T-ARMS) mutation was used to analyze the remaining SNPs. As a result, we could isolate the same group or select an individual variety using the nine unique SNPs from SNP780 to SNP9183. Furthermore, nucleotide sequence analysis for the region confirmed that the alleles were identical. In conclusion, our results show that combining SNP analysis of the whole silkworm genome with the decision tree is of high value as a discriminative marker for classifying silkworm varieties.

Isolation of Agarivorans sp. KC-1 and Characterization of Its Thermotolerant β-Agarase (한천분해세균 Agarivorans sp. KC-1의 분리 및 내열성 β-아가라제의 특성 규명)

  • Min, Kyung-Cheol;Lee, Chang-Eun;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1056-1061
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    • 2018
  • This article reports an agar-degrading marine bacterium and characterizes its agarase. The agar-degrading marine bacterium, KC-1, was isolated from seawater on the shores of Sacheon, in Gyeongnam province, Korea, using Marine Broth 2216 agar medium. To identify the agar-degrading bacterium as Agarivorans sp. KC-1, phylogenetic analysis based on the 16S rRNA gene sequence was used. An extracellular agarase was prepared from a culture medium of Agarivorans sp. KC-1, and used for the characterization of enzyme. The relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 65, 91, 96, 100, 77, and 35%, respectively. The relative activities at pH 5, 6, 7, and 8 were 93, 100, 87, and 82%, respectively. The extracellular agarase showed maximum activity (254 units/l) at pH 6.0 and $50^{\circ}C$ in 20 mM of Tris-HCl buffer. The agarase activity was maintained at 90% or more until 2 hr exposure at $20^{\circ}C$, $30^{\circ}C$ and $40^{\circ}C$, but it was found that the activity decreased sharply from $60^{\circ}C$. A zymogram analysis showed that Agarivorans sp. KC-1 produced 3 agar-degrading enzymes that had molecular weights of 130, 80, and 69 kDa. A thin layer chromatography analysis suggested that Agarivorans sp. KC-1 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarooligosaccharides, including neoagarohexaose (21.6%), neoagarotetraose (32.2%), and neoagarobiose (46.2%). These results suggest that Agarivorans sp. KC-1 and its thermotolerant ${\beta}$-agarase would be useful for the production of neoagarooligosaccharides that inhibit bacterial growth and delay starch degradation.