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Gene Expression According to Electromyostimulation after Atrophy Conditions and Muscle Atrophy in Skeletal Muscle

  • Park, Chang-Eun
    • Biomedical Science Letters
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    • v.18 no.1
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    • pp.49-55
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    • 2012
  • Numerous biochemical molecules have been implicated in the development of muscular atrophy. However, control mechanisms associated with muscular disease are not clear. The present study was conducted to investigate gene expression profiles of rat muscle during the denervation to atrophy transition processes. We isolated total RNA from rats suffering from partial muscle atrophy (P) and electromyostimulated atrophy (PE) and synthesized cDNA using annealing control primers. Using 20 ACPs for PCR, we cloned 18 DEGs using TOPO TA cloning vector, sequenced, and analyzed their identities using BLAST search. Sequences of 14 clones significantly matched database entries, while one clone was ESTs, and 3 clones were unidentified. Different expression profiles of selected DEGs between P and PE were confirmed. The troponin T, Fkbp1a, RGD1307554, Phtf1, Atp1a1 and Commd3 were highly expressed genes in the P and PE groups, while Krox-25 and TCOX2 were only expressed genes in the P group, the Sv2b and Marcks were only expressed genes in PE group. also, Cox8h was highly expressed genes in PE groups. The ASPH, ND1, and ARPL1 were highly expressed genes in the P and PE groups. List of genes obtained from the present study might provide an insight for the study of mechanism regulating muscle atrophy and electrostimulated muscle atrophy transitions. These data suggest that troponin T, Fkbp1a, RGD1307554, Phtf1, Atp1a1, and Commd3 are potentially useful as clinical biomarkers of age-related muscle atrophy and dysfunction.

Isolation and Identification of Yeasts from Wild Flowers Collected around Jangseong Lake in Jeollanam-do, Republic of Korea, and Characterization of the Unrecorded Yeast Bullera coprosmaensis

  • Han, Sang-Min;Hyun, Se-Hee;Lee, Hyang Burm;Lee, Hye Won;Kim, Ha-Kun;Lee, Jong-Soo
    • Mycobiology
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    • v.43 no.3
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    • pp.266-271
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    • 2015
  • Several types of yeasts were isolated from wild flowers around Jangseong Lake in Jeollanam-do, Republic of Korea and identified by comparing the nucleotide sequences of the PCR amplicons for the D1/D2 variable domain of the 26S ribosomal DNA using Basic Local Alignment Search Tool (BLAST) analysis. In total, 60 strains from 18 species were isolated, and Pseudozyma spp. (27 strains), which included Pseudozyma rugulosa (7 strains) and Pseudozyma aphidis (6 strains), was dominant species. Among the 60 strains, Bullera coprosmaensis JS00600 represented a newly recorded yeast strain in Korea, and its microbiological characteristics were investigated. The yeast cell has an oval-shaped morphology measuring $1.4{\times}1.7{\mu}m$ in size. Bullera coprosmaensis JS00600 is an asporous yeast that exhibits no pseudomycelium formation. It grew well in vitamin-free medium as well as in yeast extract-malt extract broth and yeast extract-peptone-dextrose (YPD) broth, and it is halotolerant growing in 10% NaCl-containing YPD broth.

groES Expression Related to Antifungal Activity of Streptomyces sp. SAR01 (Streptomyces sp. SAR01 균주에서의 항진균 관련 groES의 발현)

  • 이영근;김재성;조규성;장병일;추철형
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.162-167
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    • 2002
  • To analyse proteins and gene related to antifungal activity, SAR01 strain was isolated from a brown seaweed and identified as Streptomyces sp. by FAME(fatty acid methyl ester) analysis. Antifungal activity deficient mutant(SAR535) of Streptomyces sp. SAR01 was induced by gamma radiation$({60}^Co)$. It was found that 6 specific protein spots appeared only in SAR01 by 2-D electrophoresis analysis. Among them, a protein of 10 kDa had homology of 96% with 10 kD chaperonin cpn 10 (GroES) by Basic Local Alignment Search Tool(BLAST, NCBI) analysis. SAR535 transformants into which groES was transferred by electroporation revealed antifungal activity newly similar with SAR01 It suggested that groES be supposed to be related to the antifungal activity of Streptomyces sp. SAR01.

Identification of Differentially Expressed Genes by Exposure of Methylmercury in Neuroblastoma Cell Line Using Suppression Subtractive Hybridization (SSH)

  • Kim, Youn-Jung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.2 no.1
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    • pp.60-66
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    • 2006
  • Methylmercury (MeHg), one of the heavy metal compounds, can cause severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. This study, using of suppression subtractive hybridization (SSH) method, was peformed to identify differentially expressed genes by MeHg in SH-SY5Y human neuroblastoma cell line. We prepared to total RNA from SH-SY5Y cells treated with solvent (DMSO) and $6.25\;{\mu}M\;(IC_{50})$ MeHg and performed forward and reverse SSH. Differentially expressed cDNA clones were screened by dot blot, sequenced and confirmed that individual clones indeed represent differentially expressed genes with real time RT-PCR. These sequences were identified by BLAST homology search to known genes or expressed sequence tags (ESTs). Analysis of these sequences may provide an insight into the biological effects of MeHg in the pathogenesis of neurodegenerative disease and a possibility to develop more efficient and exact monitoring system of heavy metals as ubiquitous environmental pollutants.

Isolation of Three Unrecorded Yeasts from the Guts of Earthworms Collected from Korea

  • Oh, Hyejin;Kim, Myung Kyum
    • The Korean Journal of Mycology
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    • v.49 no.4
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    • pp.545-553
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    • 2021
  • In 2021, seven yeast strains were isolated from earthworm (Eisenia andrei) gut samples collected from the Nanji Water Regeneration Center in Goyang City, Gyeonggi Province, Korea. A total of seven yeasts were isolated, of which three strains have not been previously reported in Korea. To identify the yeasts, pairwise sequence comparisons of large subunit (LSU) rDNA sequences were performed using the basic local alignment search tool (BLAST). Assimilation test and cell morphology analysis were performed using the API 20C AUX kit and phase contrast microscope, respectively. Five of the seven strains were assigned to the genus Candida of the order Saccharomycetales of the class Saccharomycetes, and two to the genus Apiotrichum of the order Trichosporonales of the class Tremellomycetes. The yeast strain Candida sojae E2 belongs to the family Debaryomycetaceae, and Apiotrichum laibachii E8 and A. laibachii E9 belong to the family Trichosporonaceae. All strains were cultured in yeast mold agar for three days and showed different colony forms. C. sojae E2 was round and entire shaped, while A. laibachii E8 and A. laibachii E9 was round and convex shaped. This study focuses on the description of the three yeast strains that have not been officially reported in Korea.

De-novo Hybrid Protein Design for Biodegradation of Organophosphate Pesticides

  • Awasthi, Garima;Yadav, Ruchi;Srivastava, Prachi
    • Microbiology and Biotechnology Letters
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    • v.47 no.2
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    • pp.278-288
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    • 2019
  • In the present investigation, we attempted to design a protocol to develop a hybrid protein with better bioremediation capacity. Using in silico approaches, a Hybrid Open Reading Frame (Hybrid ORF) is developed targeting the genes of microorganisms known for degradation of organophosphates. Out of 21 genes identified through BLAST search, 8 structurally similar genes (opdA, opd, opaA, pte RO, pdeA, parC, mpd and phnE) involved in biodegradation were screened. Gene conservational analysis categorizes these organophosphates degrading 8 genes into 4 super families i.e., Metallo-dependent hydrolases, Lactamase B, MPP and TM_PBP2 superfamily. Hybrid protein structure was modeled using multi-template homology modeling (3S07_A; 99%, 1P9E_A; 98%, 2ZO9_B; 33%, 2DXL_A; 33%) by $Schr{\ddot{o}}dinger$ software suit version 10.4.018. Structural verification of protein models was done using Ramachandran plot, it was showing 96.0% residue in the favored region, which was verified using RAMPAGE. The phosphotriesterase protein was showing the highest structural similarity with hybrid protein having raw score 984. The 5 binding sites of hybrid protein were identified through binding site prediction. The docking study shows that hybrid protein potentially interacts with 10 different organophosphates. The study results indicate that the hybrid protein designed has the capability of degrading a wide range of organophosphate compounds.

Viruses Associated with Fig Mosaic Disease in Different Fig Varieties in Montenegro

  • Latinovic, Jelena;Radisek, Sebastjan;Bajceta, Milija;Jakse, Jernej;Latinovic, Nedeljko
    • The Plant Pathology Journal
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    • v.35 no.1
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    • pp.32-40
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    • 2019
  • Symptoms of fig mosaic disease have been noticed on leaves of fig (Ficus carica) for several decades, in Montenegro. In 2014, leaf samples were collected from trees of six fig cultivars in a plantation located in the main fig-producing area of Montenegro, to study the disease. After RNA isolation, samples were tested by RT-PCR for detection of nine fig viruses and three viroids. Four viruses were detected: fig leaf mottle-associated virus 1 (FLMaV-1), fig mosaic virus (FMV), fig mild mottle-associated-virus (FMMaV) and fig badnavirus 1 (FBV-1). Most of the viruses were present in mixed infections. The amplicons of the viruses were directly sequenced from both directions. A BLAST search of these sequences revealed sequence identities with their closest counterparts at GenBank of 92, 97, 92 and 100%, for FLMaV-1, FMV, FMMaV and FBV-1, respectively. Different responses in symptom expression due to the various virus combinations detected have been demonstrated. Variety $Su{\check{s}}ilica$ had the least symptom expression, with only one virus (FBV-1) found. Considering that the production of figs in Montenegro is increasing and has a substantial relevance in this geographic location, the results indicate that more attention should be given to improving the phytosanitary condition of fig trees in the country.

A New Putative Chitinase from Reticulitermes speratus KMT001

  • Ham, Youngseok;Park, Han-Saem;Kim, Yeong-Suk;Kim, Tae-Jong
    • Journal of the Korean Wood Science and Technology
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    • v.47 no.3
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    • pp.371-380
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    • 2019
  • Termites are pests that cause serious economic and cultural damage by digesting wood cellulose. Termites are arthropods and have an epidermis surrounded by a chitin layer. To maintain a healthy epidermis, termites have chitinase (${\beta}$-1,4-poly-N-acetyl glucosamidinase, EC 3.2.1.14), an enzyme that hydrolyzes the ${\beta}$-1,4 bond of chitin. In this study, the amino acid sequence of the gene, which is presumed to be termite chitinolytic enzyme (NCBI accession no. KC477099), was obtained from a transcriptomic analysis of Reticulitermes speratus KMT001 in Bukhan Mountain, Korea. An NCBI protein BLAST search confirmed that the protein is a glycoside hydrolase family 18 (GH18). The highest homology value found was 47%, with a chitinase from Araneus ventricosus. Phylogenetic analysis indicated that the KC477099 protein has the same origins as those of arthropods but has a very low similarity with other arthropod chitinases, resulting in separation at an early stage of evolution. The KC477099 protein contains two conserved motifs, which encode the general enzymatic characteristics of the GH18 group. The amino acid sequences $Asp^{156}-Trp^{157}-Glu^{158}$, which play an important role in the enzymatic activity of the GH18 group, were also present. This study suggests that the termite KC477099 protein is a new type of chitinase, which is evolutionarily distant from other insect chitinases.

Identification of chromosomal translocation causing inactivation of the gene encoding anthocyanidin synthase in white pomegranate (Punica granatum L.) and development of a molecular marker for genotypic selection of fruit colors

  • Jeong, Hyeon-ju;Park, Moon-Young;Kim, Sunggil
    • Horticulture, Environment, and Biotechnology : HEB
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    • v.59 no.6
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    • pp.857-864
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    • 2018
  • Previous studies have not detected transcripts of the gene encoding anthocyanidin synthase (ANS) in white pomegranates (Punica granatum L.) and suggest that a large-sized insertion in the coding region of the ANS gene might be the causal mutation. To elucidate the identity of the putative insertion, 3887-bp 5' and 3392-bp 3' partial sequences of the insertion site were obtained by genome walking and a gene coding for an expansin-like protein was identified in these genome-walked sequences. An identical protein (GenBank accession OWM71963) isolated from pomegranate was identified from BLAST search. Based on information of OWM71963, a 5.8-Mb scaffold sequence with genes coding for the expansin-like protein and ANS were identified. The scaffold sequence assembled from a red pomegranate cultivar also contained all genome-walked sequences. Analysis of positions and orientations of these genes and genome-walked sequences revealed that the 27,786-bp region, including the 88-bp 5' partial sequences of the ANS gene, might be translocated into an approximately 22-kb upstream region in an inverted orientation. Borders of the translocated region were confirmed by PCR amplification and sequencing. Based on the translocation mutation, a simple PCR codominant marker was developed for efficient genotyping of the ANS gene. This molecular marker could serve as a useful tool for selecting desirable plants at young seedling stages in pomegranate breeding programs.

Isolation of four unrecorded yeasts in the family Filobasidiaceae from soil in Korea

  • Maeng, Soohyun;Park, Yuna;Srinivasan, Sathiyaraj
    • Journal of Species Research
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    • v.10 no.4
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    • pp.350-355
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    • 2021
  • In 2020, 11 Basidiomycetous yeast strains were isolated from soil samples collected from the forests of Namhansanseong in Korea. Among them, seven species were reported, but four species were unreported in Korea. To identify wild yeasts, pairwise sequence comparisons of D1/D2 domain of the 26S rRNA were performed using Basic Local Alignment Search Tool (BLAST). The cell morphologies and assimilation test are observed by phase contrast microscope and API 20C AUX kit, respectively. The 11 strains were assigned to the genera Rhodotorula (4 strains) of the order Sporidiobolales of the class Microbotryomycetes; and Cryptococcus(2 strains), Goffeauzyma (1 strains), Naganishia (2 strains) of the order Filobasidiales and Saitozyma (2 strains) of the order Tremellales of the class Tremellomycetes in the phylum Basidiomycota. The unreported yeast strains Cryptococcus gastricus 20n5-2, Goffeauzyma gilvescens 20n2-7, Naganishia adeliensis 20n8-1, and Naganishia friedmannii 20n24-1 belong to the family Filobasidiaceae. All strains had oval shaped cells and cream-colored colonies cultured on on YM agar for 3 days. In this study, we focus on the description of four unreported yeast species in Korea.