• Title/Summary/Keyword: Molecular Characteristics

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Characterization of Mutations in AlHK1 Gene from Alternaria longipes: Implication of Limited Function of Two-Component Histidine Kinase on Conferring Dicarboximide Resistance

  • Luo, Yiyong;Yang, Jinkui;Zhu, Mingliang;Yan, Jinping;Mo, Minghe;Zhang, Keqin
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.15-22
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    • 2008
  • Four series (S, M, R, and W) of Alternaria longipes isolates were obtained based on consecutive selection with Dimethachlon (Dim) and ultraviolet irradiation. These isolates were then characterized according to their tolerance to Dim, sensitivity to osmotic stress, and phenotypic properties. All the selected Dim-resistant isolates showed a higher osmosensitivity than the parental strains, and the last generation was more resistant than the first generation in the M, R, and W series. In addition, the changes in the Dim resistance and osmotic sensitivity were not found to be directly correlated, and no distinct morphologic characteristics were found among the resistant and sensitive isolates, with the exception of the resistant isolate K-11. Thus, to investigate the molecular basis of the fungicide resistance, a group III two-component histidine kinase (HK) gene, AlHK1, was cloned from nineteen A. longipes isolates. AlHK1p was found to be comprised of a six 92-amino-acid repeat domain (AARD), HK domain, and response regulator domain, similar to the Os-1p from Neurospora crassa. A comparison of the nucleotide sequences of the AlHK1 gene from the Dim-sensitive and -resistant isolates revealed that all the resistant isolates contained a single-point mutation in the AARD of AlHK1p, with the exception of isolate K-11, where the AlHK1p contained a deletion of 107 amino acids. Moreover, the AlHK1p mutations in the isolates of each respective series involved the same amino acid substitution at the same site, although the resistance levels differed significantly in each series. Therefore, these findings suggested that a mutation in the AARD of AlHK1p was not the sole factor responsible for A. longipes resistance to dicarboximide fungicides.

Peptide Profiling and Selection of Specific-Expressed Peptides in Hypoglycemic Sorghum Seed using SELDI-TOF MS (SELDI-TOF MS를 활용한 혈당강하 수수 종자의 펩타이드 프로파일링 및 특이 발현 펩타이드 선발)

  • Park, Sei Joon;Hwang, Su Min;Park, Jun Young;Ko, Jee-Yeon;Kim, Tae Wan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.3
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    • pp.252-262
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    • 2014
  • Sorghum seed is traditionally used as secondary food sources in addition to rice in Korea. While the hypoglycemia regulating phytochemicals have been found in sorghum seed, peptides related with hypoglycemia never been studied before. To obtain the peptide characteristics and the specifically high-expressed peptides in hypoglycemic sorghum seed, peptide profiles of seven hypoglycemic and five non-hypoglycemic sorghum lines bred in RDA were determined using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS). The twelve sorghum lines exhibited 104 peptides on CM10 protein chip array (weak cation exchange) and 95 peptides on Q10 (weak cation exchange) in the molecular mass range from 2,000 to 20,000 Da. Heat map via supervised hierarchical clustering of the significantly different peptides (p < 0.01) in peak intensity among the 12 lines effectively revealed the specifically upregulated peptides in each line and distinguished between 7 hypoglycemic and 5 non-hypoglycemic lines. Through the comparison with hypoglycemic and non-hypoglycemic lines, 10 peptides including 2231.6, 2845.4, 2907.9, 3063.5, 3132.6, 3520.8, 4078.8, 5066.2, 5296.5, 5375.5 Da were specifically high-expressed in hypoglycemic lines at p < 0.00001. This study characterized seed peptides of 12 sorghums and found ten peptides highly expressed for hypoglycemic sorghum lines, which could be used as peptide biomarkers for identification of hypoglycemic sorghum.

Isolation and Characterization of Exopolysaccharide Producing Lactic Acid Bacteria from Kimchi (김치로부터 Exopolysaccharide 생성 유산균의 분리 및 특성 규명)

  • Kim Uyo-Ju;Chang Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.196-203
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    • 2006
  • Three slime-forming lactic acid bacteria were isolated from Kimchi and shown to produce viscous exopolysaccharides (EPS) in sucrose media. The isolated strains, GJ2, C3 and C11, were identified as Leuconostoc kimchii, Leuconostoc citreum and Leuconostoc mesenteroides, respectively, by examining their metabolic characteristics and determining their 16S rDNA sequences. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 exhibited high viability (maintained initial viable cell count of $10^8$ CFU/ml) in 0.05 M sodium phosphate buffer (pH 3.0) for 2 h, in artificial gastric juice for 2 h and in 0.3% oxgall for 24 h. When tested, Leu. kimchii GJ2, in particular, displayed antimicrobial activity against pathogenic microorganisms. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 produced 21.49 g/l, 16.46 g/l and 22.98 g/l EPS, respectively, in sucrose (5%) medium. The amount of purified EPS extracted from Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 was 14.61 g/l, 7.73 g/l and 4.77 g/l, respectively. Although the EPS produced by Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 differed in viscosity, TLC and HPLC analysis revealed that each contained only one type of monosaccharide, glucose. The average molecular mass of EPS produced by Leu. kimchii GJ2 was 306,606 Da.

An Ecological Analysis of Lichens Distributed in Rocks of Coast and Field in U-do by Molecular Technique (분자생물학적 기법에 의한 우도해안과 노지암석에 분포하는 지의류의 생태학적 분석)

  • Gang, Hyeong-Il;Yun, Byeong-Jun;Kim, Seong-Hyeon;Sin, Deok-Ja;Kim, Hyeon-U;Heo, Jae-Seon;Gang, Ui-Seong;O, Gye-Heon;Go, Yeong-Jin
    • Microbiology and Biotechnology Letters
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    • v.32 no.4
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    • pp.334-340
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    • 2004
  • This study was initially performed to obtain fundamental data on lichen distribution depending on environmental condiLichens distributed on coastal rocks atleast 20 m far away from the coast were also investigated. Analyof morphological characteristics and rDNA ITS clones revealed that lichens containing 9 families and 15 genera were distributed in coastal rocks of V-do, while lichens containing 10 families and 14 genera were in field rocks, demonstrating little difference of diversity between both sites. Foliose lichens such as PhaeophysPhyscia, Pyxine belonging to the family Physciaceae were representatives distributed in coastal rocks of V-do, along with Xanthoparmelia belonging to Parmeliaceae, fruticose lichens such as Ramalina to Ramaliand crustose lichen Lecanora to Lecanoraceae. In contrast, foliose lichen Cladonia belonging to the family Cladoniaceae, fruitcose lichen Stereocaulon to Streocaulaceae, and crustose lichen Porpidia to Porpidwere found only on the surface of field rocks. Crustose lichens containing genera Caloplaca, CanDirinaria, Graphis, Rhizocarpon, and Pertusaria were rarely distributed in coastal rocks, and most of them were also found in field rocks.

Pink Mold Rot on Unishiu Orange (Citrus unshiu Mac.) Caused by Trichothecium roseum (Pers.) Link ex Gray in Korea (Trichothecium roseum에 의한 감귤 분홍빛열매썩음병 발생)

  • Kwon, Jin-Hyeuk;Kang, Dong-Wan;Choi, Okhee;Shim, Hong-Sik
    • Research in Plant Disease
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    • v.19 no.3
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    • pp.226-228
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    • 2013
  • In 2012, a pink mold rot was observed on unishiu orange (Citrus unshiu Mac.) fruits at the Wholesale Market for Agricultural Products, Jinju, Korea. The symptom on unishiu orange was a water-soaked lesion on the surface of fruit, which later on enlarged to form softened brown rot lesions. The diseased fruits were covered with pink-colored mold, consisting of conidia and conidiophores of the pathogen. Optimum temperature for mycelial growth was $25^{\circ}C$. Conidia were hyaline, smooth, 2-celled, and thick-walled conidia with truncate bases, ellipsoidal to pyriform, characteristically held together zig-zag chains and $12-26{\times}8-12{\mu}m$ in size. Conidiophore was erect, colorless, unbranched, and 4-5 ${\mu}m$ wide. On the basis of mycological characteristics, pathogenicity test, and molecular analysis with complete ITS rDNA region, the causal fungus was identified as Trichothecium roseum (Pers.) Link ex Gray. This is the first report of pink mold rot caused by T. roseum on unishiu orange in Korea.

Selection and Agronomic Traits of Radiation-induced Variants in Rice (방사선 이용 벼 돌연변이 계통 선발 및 농경 형질조사)

  • Lee, In-Sok;Kim, Dong-Sub;Lee, Sang-Jae;Song, Hi-Sub;Lim, Yong-Pyo;Lee, Young-Il
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.19-25
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    • 2003
  • Radiation technique in agriculture was initiated to develop mutant rice. Seeds of Daechungbyeo rice were irradiated with 250 Gy gamma ray for the purpose of inducing and selecting rice variants. Some quantitative traits of the variants in M$_{8}$ generation were evaluated and RAPD analysis was carried out. Variants showed a wider range of agronomic characteristics in both a positive and a negative direction compared with their original variety. The new mutants were characterized by an increased or decreased in plant height, lodging resistance and shorter panicle. RAPD analysis showed that polymorphic bands were presented in most of the primers. In comparison with the original variety, variants were classified into four groups through UPGMA analysis. Among mutants no. 91, 139, 140 and 141 was ranked as salt tolerance and the proline content of these mutants was more increased than that of original variety. The lines of 139, 140 and 141 had the highest genetic distance as compared to original variety in the dendrogram. It is expected that such variants will be useful not only for studying molecular genetics but also for breeding research and genetic analysis.s.

AGL gene mutation and clinical features in Korean patients with glycogen storage disease type III (한국인 제3형 당원병 환자의 임상상 및 AGL 유전자형)

  • Ko, Jung Min;Lee, Jung Hyun;Kim, Gu-Hwan;Yoo, Han-Wook
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.6 no.1
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    • pp.15-23
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    • 2006
  • Purpose: Glycogen storage disease type III (GSD-III), is a rare autosomal recessive disorder of glycogen metabolism. The affected enzyme is amylo-1,6-glucosidase, 4-alpha-glucanotransferase (AGL, glycogen debranching enzyme), which is responsible for the debranching of the glycogen molecule during catabolism. The disease has been demonstrated to show clinical and biochemical heterogeneity, reflecting the genotype-phenotype heterogeneity among different patients. In this study, we analyzed mutations of the AGL gene in three unrelated Korean GSD-III patients and discussed their clinical and laboratory implications. Methods: We studied three GSD-III patients and the clinical features were characterized. Sequence analysis of 35exons and part exon-intron boundaries of the AGLgene in patients were carried out by direct DNA sequencing method using genomic DNA isolated from patients' peripheral leukocytes. Results: The clinical features included hepatomegaly (in all patients), seizures (in patient 2), growth failure (in patients 1), hyperlipidemia (in patients 1 and 3), raised transaminases and creatinine kinase concentrations (in all patients) and mild EKG abnormalities (in patients 2). Liver transplantation was performed in patient 2due to progressive hepatic fibrosis. Administration of raw-corn-starch could maintain normoglycemia and improve the condition. DNA sequence analysis revealed mutations in 5 out of 6 alleles. Patient 1 was a compound heterozygote of c.1282 G>A (p.R428K) and c.1306delA (p.S603PfsX6), patient 2 with c.1510_1511insT (p.Y504LfsX10), and patient 3 with c.3416 T>C (p.L1139P) and c.l735+1 G>T (Y538_R578delfsX4) mutations. Except R428K mutation, 4 other mutations identified in3 patients were novel. Conclusion: GSD-III patients have variable phenotypic characteristics resembling GSD-Ia. The molecular defects in the AGL gene of Korean GSD-III patients were genetically heterogeneous.

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Characteristics of Gentamicin Resistant Pseudomonas aeruginosa (녹농균의 항생제 내성의 특성)

  • Kim, Sang-Yoon;Lee, Yoo-Chul;Seol, Sung-Yong;Cho, Dong-Taek;Chun, Do-Ki
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.1-16
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    • 1986
  • Fifty-one strains of Pseudomonas aeruginosa were isolated from various clinical specimens. Among them, 26 (51%) strains were gentamicin-resistant (Gm') and 25 (49%) were susceptible to gentamicin (Gm'). The frequencies of resistant strains to piperacillin (Pi), cefotaxime, moxalactam, cefoperazone (Cz), and amikacin (Ak) ranged from 21.6 to 31.4%, and $MIC_{50}$ of these drugs were lower than the critical concentrations of susceptibility and resistance. Thirty (58.8%) strains were multiply resistant to 12 or more drugs. All Gm' strains were multiply resistant to 12 or more drugs and one was resistant to all 18 drugs tested, while only four Gm' strains were multiply resistant to 12 drugs and the multiplicity of resistance of the other Gm' strains were less than 10 drugs. Resistance to Gm appeared to have a significant correlation with the resistance to tobramycin (Tb), Ak, Pi, and Cz. All Gm' strains were resistant to Tb and about 38.4 to 46.1% of them were resistant to Ak, Pi, and Cz. The incorporation of $Ca^{++}$ and $Mg^{++}$ ions in Mueller-Hinton agar (MHA) did not influence the MICs of Gm, Tb, carbenicillin (Cb), Pi, and Cz as compared with the results obtained in MHA without these ions. Gm strains were studied on the combined effect of beta-lactam antibiotics and aminoglycosides by the methods of checkerboard and modified paper strip diffusion. Most Gm' strains showed significant synergistic effects by the FIC index between Ak and three beta-lactam antibiotics; Cb, Pi, and Cz, but these results did not in agreement the results obtained through the method of modified paper strip diffusion test. In order to know the nature of the drug resistance of P. aernginosa, the plasmid profile analysis was studied. Agarose gel electrophoresis of lysates processed by the method of Kado and Liu showed one or more plasmids in 22 (43.1%) strains. A group of 19 strains showed at least one band of plasmid and three strains two bands. The range of the molecular weight of plasmids was 3.8 to 243 Mdal. All strains carrying large plasmids larger than 200 Mdal were isolated from wound specimens. Three Gm' strains also harboured the plasm ids of 13 to 203 Mdal.

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Purification and Characterization of $L-galactono-{\gamma}-lactone$ Oxidase in Pichia sp. Isolated from Kimchi (김치유래 Pichia속 효모가 생산하는 $L-galactono-{\gamma}-lactone\;oxidase$의 분리 정제 및 특성)

  • Oh, Ji-Young;Han, Young-Sook
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1135-1142
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    • 2003
  • The purification and characteristics of the biosynthesis enzyme of vitamin C from microorganisms related with kimchi fermentation were investigated to define vitamin C biosynthetic pathways in yeast. A yeast strain (Pichia onychis 16-4) which synthesizes vitamin C with galacturonic acid as substrate at high rate was isolated from kimchi. The enzyme $L-galactono-{\gamma}-lactone$ oxidase isolated from the yeast was purified and characterized. The specific activity of the crude enzyme was 7.26 unit/mg protein, which increased to 4,698 unit/mg protein with a chromatography of Sephacryl S-200HR; indicating a 647.1-fold level of purification. The molecular weights of the dissociated enzymes were estimated to be 31,000, 39,000, and 50,000 KD. Among the substrates tested, $L-galactono-{\gamma}-lactone$ was the most effective. The enzyme showed optimum activity ah pH 7.8 and 35c. The purified enzyme uses $O_2$ as the electron acceptor for oxidation of $L-galactono-{\gamma}-lactone$.

Physicochemical Properties of Depolymerized Barley β-Glucan by Alkali Hydrolysis (알칼리 가수분해에 따른 보리 β-Glucan의 이화학적 특성)

  • Lee, Sang-Hoon;Jang, Gwi-Yeong;Kim, Kee-Jong;Lee, Mi-Ja;Kim, Tae-Jip;Lee, Jun-Soo;Jeong, Heon-Sang
    • The Korean Journal of Food And Nutrition
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    • v.26 no.3
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    • pp.601-607
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    • 2013
  • This study was performed to investigate the changes of total and soluble ${\beta}$-glucan contents, purity and physicochemical characteristics of alkali hydrolyzed barley varieties: Saessalbori (SSB), Saechalssalbori (SCSB) and Hinchalssalbori (HCSB). The barleys were hydrolyzed at different concentrations of sodium hydroxide (0.2~1.0 N) for 12 hours. Total ${\beta}$-glucan contents of raw SSB, SCSB and HCSB were 8.40, 7.77 and 8.28%, and soluble ${\beta}$-glucan contents were 4.80, 4.16 and 4.61%, respectively. The total ${\beta}$-glucan contents after alkali hydrolyzed at 1.0 N NaOH were 7.54, 6.89 and 7.54%, also soluble ${\beta}$-glucan contents were 4.82, 4.30 and 4.55%, respectively. The degree of purity of soluble ${\beta}$-glucan in SSB, SCSB, and HCSB were 35.79, 30.91 and 33.90%, respectively. They were increased to 74.02, 75.28 and 81.41% after hydrolyzed at 1.0 N NaOH, respectively. The molecular weight and viscosity of soluble ${\beta}$-glucan solutions were decreased as sodium hydroxide concentration was increased. The re-solubility of raw barley ${\beta}$-glucan was about 50%; however, it was increased to approximately 87% as sodium hydroxide concentration was increased.