• Korean Journal of Microbiology
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• v.44 no.4
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• pp.311-316
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• 2008

In this study, biological PCE degradation by using a BTEX degrading bacterium, named BJ10, under aerobic conditions in the presence of toluene was examined. According to morphological, physiological characteristics, 16S rDNA sequencing and fatty acid analysis, BJ10 was classified as Pseudomonas putida. As a result of biological PCE degradation at low PCE concentrations (5 mg/L), PCE removal efficiency by P. putida BJ10 was 52.8% for 10 days, and PCE removal rate was 5.9 nmol/hr (toluene concentration 50 mg/L, initial cell density 1.0 g (wet weight)/L, temperature 30, pH 7 and DO $3.0{\sim}4.2\;mg/L$. At high PCE concentration (100 mg/L), PCE removal efficiency by P. putida BJ10 was 20.3% for 10 days, and PCE removal rate was 46.0 nmol/hr under the same conditions. The effects of various toluene concentration (5, 25, 50, 100, 200 mg/L) on PCE degradation were examined under the same incubation conditions. The highest PCE removal efficiency of PCE was 57.0% in the initial PCE concentration of 10 mg/L in the presence of 200 mg/L toluene for 10 days. Furthermore, the additional injection of 5.5 mg/L PCE (total 7.6 mg/L) made 63.0% degradation for 8 days in the presence of 50 mg/L toluene under the same conditions. Its removal rate was 13.5 nmol/hr, which was better than the initial removal rate (8.1 nmol/hr).

• Journal of Embryo Transfer
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• v.29 no.4
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• pp.339-344
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• 2014

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas9) system can be applied to produce transgenic pigs. Therefore, we applied CRISPR/Cas9 system to generate FoxN1-targeted pig parthenogenetic embryos. Using single guided RNA targeted to pig FoxN1 genes was injected into cytoplasm of in vitro matured oocyte before electrical activation. In results, regardless of the concentrations of vector, the cleavage rate were significantly (p<0.05) decreased ($4ng/{\mu}l$, 51.24%; $8ng/{\mu}l$, 40.88%; and $16ng/{\mu}l$; 45.22%) compared to no injection group (70.44%). The blastocyst formation rates were also decreased in vector injected 3 groups ($4ng/{\mu}l$, 7.96%; $8ng/{\mu}l$, 6.4%; and $16ng/{\mu}l$; 9.04%) compared to no injection group (29.07%). In addition, the blastocyst formation rates between sham injected group (13.51%) and no injection group (29.07%) also showed significant difference (p<0.05). The mutation rates were comparable between groups ($4ng/{\mu}l$, 18.4%; $8ng/{\mu}l$, 12.5%; and $16ng/{\mu}l$; 20.0%). The sequencing analysis showed that blastocysts derived from each group were successfully mutated in FoxN1 loci regardless of the vector concentrations. However, the deletion patterns were higher than the patterns of point mutation and insertion regardless of the vector concentrations. In conclusion, we described that cytoplasmic microinjection of FoxN1-targeted CRISPR/Cas9 vector could efficiently generate transgenic pig parthenogenetic embryos in one-step.

• The Korean Journal of Mycology
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• v.38 no.2
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• pp.160-166
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• 2010

A homothallic filamentous fungus Aspergillus nidulans has been used as the a model organism for studying growth and development for eukaryotic system. Various studies about specific transcription factors have been performed for elucidating the molecular mechanisms of growth, asexual and sexual developmental processes. Among them, the fkhE gene (AN2025.3) is located in chromosome VII and contains an ORF encoding 718 amino acid polypeptide intervening with two short introns. The cDNA sequencing revealed that at least four types of alternative splicing events were occurred when the fkhE gene was transcribed. The putative FkhE polypeptide contains a conserved forkhead domain and a bipartite nuclear localization signal at it's N-terminus and C-terminus, respectively. Deletion of fkhE resulted in impaired conidiophore formation in a solid medium. However, the sexual developmental process or cleistothecia formation was normal. Furthermore, fkhE deletion mutant produced conidiophores and conidia under the submerged culture, indicating that the fkhE gene is involved in asexual developmental process similar to the fkhF gene.

• Korean Journal of Microbiology
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• v.47 no.4
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• pp.281-288
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• 2011

To screen downstream genes of Aspergillus nidulans MsnA showing amino acid sequence similarity to the zinc finger region of Msn2/4 stress response transcription factors in Saccharomyces cerevisiae, differentially expressed genes (DEG) in MsnA overexpressed or msnA null mutant strains compared to wild type have been isolated. The cognate gene IDs were identified by DNA sequencing of the selected DEGs. Among those, DEG6 was known as mstB encoding a putative monosaccharide transporter. Expression level of mstB mRNA was increased in MsnA overproducing strains and MsnA bound directly to the promoter region of mstB in vitro. MstB containing twelve transmembrane domains exhibited 80% of amino acid sequence identities to A. niger MstA a high-affinity monosaccharide transporter. A null mutant of mstB was phenotypically undistinguishable to wild type. On the other hand, forced overexpression of MstB caused the increased formation of sexual structure cleistothecia in 0.1% glucose condition where wild type showed almost no cleistothecia. This result implies that mstB is involved in transport of monosaccharide required for sexual differentiation.

• Journal of Chitin and Chitosan
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• v.23 no.4
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• pp.285-292
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• 2018

In this study, for the purpose of decomposing food waste, the strain was screened from traditional fermented food and soils. The enzyme activity (protease, amylase, cellulase, lipase) experiment was carried out using the paper disc method in 212 strains isolated from 5% NaCl media. Among them, only the strains having enzyme activity of more than 2 (soil) or more than 4 (traditional fermented food) with the halozone of enzyme activity of 15 mm or more were selected first, and microorganism identification through 16S rRNA sequencing was performed. Finally, were identified such as Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus siamensis, Bacillus licheniformis, Bacillus aquimaris, Bacillus megaterium, Bacillus koreensis, Bacillus stratoshericus, Bacillus aryabhattai, Bacillus safensis, Marinobacter hydrocarbonoclasticus. 11 species of mixed strains were confirmed that the culture time was 24 hours, the incubation temperature was $30^{\circ}C$ and the optimum pH was 7.0. In order to confirm the degree of decomposition of standard food wastes (100 g) by treating 11 kinds of mixed strains (25%), solid content of more than $2000{\mu}m$ was determined to be 103 g for the sterilized water group and 18 g for the mixed strains group. And the rest was decomposed to a size of less than $2000{\mu}m$.

• Journal of Biomedical and Translational Research
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• v.19 no.4
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• pp.116-124
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• 2018

Korean Native Pig (KNP) has a uniform black coat color, excellent meat quality, white colored fat, solid fat structure and good marbling. However, its growth performance is low, while the western origin Yorkshire pig has high growth performance. To take advantage of the unique performance of the two pig breeds, we raised crossbreeds (KNP ${\times}$ Yorkshire to make use of the heterotic effect. We then analyzed the liver transcriptome as it plays an important role in fat metabolism. We sampled at two stages: 10 weeks and at 26 weeks. The stages were chosen to correspond to the change in feeding system. A total of 16 pigs (8 from each stage) were sampled and RNA sequencing was performed. The reads were mapped to the reference genome and differential expression analysis was performed with edgeR package. A total of 324 genes were found to be significantly differentially expressed (${\left|log2FC\right|}$ > 1 & q < 0.01), out of which 180 genes were up-regulated and 144 genes were down-regulated. Principal Component Analysis (PCA) showed that the samples clustered according to stages. Functional annotation of significant DEGs (differentially expressed genes) showed that GO terms such as DNA replication, cell division, protein phosphorylation, regulation of signal transduction by p53 class mediator, ribosome, focal adhesion, DNA helicase activity, protein kinase activity etc. were enriched. KEGG pathway analysis showed that the DEGs functioned in cell cycle, Ras signaling pathway, p53 signaling pathway, MAPK signaling pathway etc. Twenty-nine transcripts were also part of the DEGs, these were predominantly Cys2His2-like fold group (C2H2) family of zinc fingers. A protein-protein interaction (PPI) network analysis showed that there were three highly interconnected clusters, suggesting an enrichment of genes with similar biological function. This study presents the first report of liver tissue specific gene regulation in a cross-bred Korean pig.

• Journal of Microbiology and Biotechnology
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• v.29 no.10
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• pp.1644-1655
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• 2019

Saccharomyces cerevisiae (S. cerevisiae) and glucagon-like peptide-2 (GLP-2) have been employed to improve the intestinal development of weaned animals. The goal of this study was to determine whether either exogenous S. cerevisiae or GLP-2 elicits major effects on fecal microbiotas and cytokine responses in weaned piglets. Ninety-six piglets weaned at 26 days were assigned to one of four groups: 1) Basal diet (Control), 2) empty vector-harboring S. cerevisiae (EV-SC), 3) GLP-2-expressing S. cerevisiae (GLP2-SC), and 4) recombinant human GLP-2 (rh-GLP2). At the start of the post-weaning period (day 0), and at day 28, fecal samples were collected to assess the bacterial communities via sequencing the V1-V2 region of the 16S-rRNA gene, and piglets' blood was also sampled to measure cytokine responses (i.e., IL-$1{\beta}$, TNF-${\alpha}$, and IFN-${\gamma}$). This study revealed that, on the one hand, although S. cerevisiae supplementation did not significantly alter the growth of weaned piglets, it induced increases in the relative abundances of two core genera (Ruminococcaceae_norank and Erysipelotrichaceae_norank) and decreases in the relative abundances of two other core genera (Lachnospiraceae_norank and Clostridiale_norank) and cytokine levels (IL-$1{\beta}$ and TNF-${\alpha}$) (p < 0.05, Control vs EV-SC; p < 0.05, rh-GLP2 vs GLP2-SC). On the other hand, GLP-2 supplementation had no significant influence on fecal bacterial communities and cytokine levels, but it produced better body weight and average daily gain (p < 0.05, Control vs EV-SC; p < 0.05, rh-GLP2 vs GLP2-SC). Therefore, altered fecal microbiotas and cytokine response effects in weaned piglets were due to S. cerevisiae rather than GLP-2.

• The Korean Journal of Malacology
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• v.29 no.3
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• pp.171-179
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• 2013

Arginine kinase (ArK) is known to play an important role in most invertebrates the level of ATP by phosphorylation of phosphagens in cell and immuninty in living organisms. ArK has been identified in many kinds of organisms ranging from invertebrate to vertebrate. However, no ArK gene has been cloned and investigated from N. samarangae. This leads us to identify ArK cDNA (NsArK) from the expressed sequence tag (EST) sequencing of N. samarangae. Sequence analysis indicated that the coding region of 1,065 bp contains 355 amino acid residues. Molecular phylogenetic analysis shows that NsArK had very high similarities with mollusca and arthropoda. In an attempt to investigate a potential role of NsArK in the digestive gland of N. samarangae, expression patterns were analyzed. RT-PCR analsysis shows that NsArK mRNA is induced in the rane of 1.2 fold at 6 hr by laminarin when compared with the control. The immunnologial and physiological role of NsArK remains to be further investigated in N. samarangae.

• Food Science and Preservation
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• v.18 no.6
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• pp.973-979
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• 2011

The grape cultivar Campbell Early has high levels of malic acid as well as tartaric acid. The high concentration of total acid in the Campbell Early wine is a critical aspect of the wine's sensory characteristics. To prevent the deterioration of the wine's quality, which is caused by the strong sour taste derived from the raw material in wine making, the deacidification factor was investigated via carbonic maceration under different temperature conditions, especially in the presence or absence of malolactic bacteria. Based on the results of the presence test of the malolactic bacteria during carbonic-maceration treatment, Lactobacillus brevis, Lactobacillus plantarum, and Streptococcus thermophilus were characterized morphologically and were identified via biochemical tests and 16S-rRNA-gene-sequencing analysis. The isolated strains were found not to consume malic acid and to produce lactic acid. Moreover, these strains were consumed as soluble solids. The isolated strains are popularly known as lactic-acid bacteria and should have produced lactic acid from glucose. The Oenococcus oeni of the malolactic bacteria was not isolated. These results showed that the isolated strains are not deacidified during carbonic-maceration treatment.

• Journal of Animal Science and Technology
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• v.63 no.4
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• pp.934-953
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• 2021

Ciglitazone is a member of the thiazolidinedione family, and specifically binds to peroxisome proliferator-activated receptor-γ (PPARγ), thereby promoting adipocyte differentiation. We hypothesized that ciglitazone as a PPARγ ligand in the absence of an adipocyte differentiation cocktail would increase adiponectin and adipogenic gene expression in bovine satellite cells (BSC). Muscle-derived BSCs were isolated from six, 18-month-old Yanbian Yellow Cattle. The BSC were cultured for 96 h in differentiation medium containing 5 µM ciglitazone (CL), 10 µM ciglitazone (CM), or 20 µM ciglitazone (CH). Control (CON) BSC were cultured only in a differentiation medium (containing 2% horse serum). The presence of myogenin, desmin, and paired box 7 (Pax7) proteins was confirmed in the BSC by immunofluorescence staining. The CL, CM, and CH treatments produced higher concentrations of triacylglycerol and lipid droplet accumulation in myotubes than those of the CON treatment. Ciglitazone treatments significantly increased the relative expression of PPARγ, CCAAT/enhancer-binding protein alpha (C/EBPα), C/EBPβ, fatty acid synthase, stearoyl-CoA desaturase, and perilipin 2. Ciglitazone treatments increased gene expression of Pax3 and Pax7 and decreased expression of myogenic differentiation-1, myogenin, myogenic regulatory factor-5, and myogenin-4 (p < 0.01). Adiponectin concentration caused by ciglitazone treatments was significantly greater than CON (p < 0.01). RNA sequencing showed that 281 differentially expressed genes (DEGs) were found in the treatments of ciglitazone. DEGs gene ontology (GO) analysis showed that the top 10 GO enrichment significantly changed the biological processes such as protein trimerization, negative regulation of cell proliferation, adipocytes differentiation, and cellular response to external stimulus. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that DEGs were involved in the p53 signaling pathway, PPAR signaling pathway, biosynthesis of amino acids, tumor necrosis factor signaling pathway, non-alcoholic fatty liver disease, PI3K-Akt signaling pathway, and Wnt signaling pathway. These results indicate that ciglitazone acts as PPARγ agonist, effectively increases the adiponectin concentration and adipogenic gene expression, and stimulates the conversion of BSC to adipocyte-like cells in the absence of adipocyte differentiation cocktail.