• Journal of the Korea Organic Resources Recycling Association
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• v.16 no.4
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• pp.64-73
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• 2008

The cDNA of CCF (coelomic cytolytic factor)-like gene (EC 3.2.1.16), a kind of glycosyl hydorlase, was isolated and cloned from the midgut of the earthworm Eisenia anderi. The size of nucleotide sequence appeared to be 1,152 bp and its predicted coding region was composed of 384 amino acid residues including the initiation methionine. The 17 residues at N-terminal end in the deduced amino acid sequence were regarded to be a signal peptide. Based on the amino acid sequence analysis, it appeared that this CCF-like protein could belong to glycosyl hydrolase family 16 (GHF16) and showed a high sequence homology of about 79~99% with CCF and CCF-like proteins from other earthworm species. The CCFs and CCF-like proteins from various earthworm species exhibited a 100% homology in the polysacchride-binding motif and glucanase motif. It has been reported that the CCFs isolated from E. fedita appeared to show a broader pattern recognition specificity than those from other earthworm species because this species resides in decaying organic matter showing very high microbial activity, implying that CCF-like protein isolated in this study from E. andrei might exhibit a broad substrate specificity that is a useful characteristic for industrial application. A phylogenetic analysis using the deduced amino acid sequences of CCF-related proteins through the BLASTX revealed that GHF16 families could be divided into three groups of metazoa, viriplantae and eubacteria subfamily. Subsequently the CCF-related proteins of metazoa subfamily could clearly be subgroup into lophotrochozoan and edysozoan type including a deuterostome origin. Further understanding of the biological properties of E. andrei CCF-like protein should be addressed to regulate the ${\beta}$-D-glucan hydrolysis and production for the industrial uses.

• Journal of Life Science
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• v.16 no.4
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• pp.626-631
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• 2006

This study was carried out to investigate the physical characteristics (height, body lenght, chest depth, head type, ear type, body color, eye type and tail type) and genetic diversity using 15 microsatellite DNA markers (PEZ 1, 5, 8, 10, 11, 12, 13, 15, 16, 17, 20, 21, FHC 2010, FHC 2054 and FHC 2079) in 44 random Miryang native dogs(6 months${\sim}$12 years old). The height, body lenght, and chest depth of Miryang native dogs were 43-55 cm(mean 49.5 cm), 45-60 cm(mean 54.3 cm), and 50-64 cm(mean 57.9 cm), respectively. Miryang native dog was medium sized. The head and eye type were reverse-triangle(100%), triangle (90.9%) and newborn moon(9.1%), respectively. Most of body color had white coat color(93.2%), light pink tongue color(100%), light black anal color(90,9%) and pink claw color(100%). The ear type showed erect ear(100%), and half-curled(56.8%), upward(34.1%), curled(9.1%) in tail type, respectively. Number of alleles observed at a single locus ranged from 2 (PEZ 21 and FHC 2010) to 14 (PEZ 13), with average number of alleles per locus of 6.13. The expected heterozygosities of 15 microsatellite loci were estimated based on gene frequencies. The highest expected heterozygosity, 0.863 was estimated in PEZ 13 locus and the lowest, 0.455 in PEZ 21 and FHC 2010 locus. And the mean expected heterozygosity of 15 microsatellite markers was calculated as 0.635. Polymorphic information content (PIC) values were ranged from 0.348 (PEZ 21 and FHC 2010) to 0.837 (PEZ 13), and the mean PIC value was calculated as 0.570. Of the 15 markers, PEZ 10, PEZ 13, PEZ 17 and FHC 2054 loci have relatively high PIC value (> 0.7) in Miryang native dog. In order to determine the efficieney of parentage control, exclusion probabilities (PE) were calculated for each allele. The highest PE 1 and PE 2 in PEZ 13 locus was caculated to 0.548 and 0.710, respectively. And the total exclusion power in PE 1 and PE 2 was calculated to 0.9895 and 0.9996, respectively. These results can give basic information for perservation and research in Miryang native dog, and phylogenetic relationships of the Korean native dog and Asian dog breeds.

• Journal of Life Science
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• v.22 no.10
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• pp.1295-1306
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• 2012

A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus atrophaeus. This species was designated as B. atrophaeus LBH-18 based on its evolutionary distance and the phylogenetic tree resulting from 16S rDNA sequencing and the neighbor-joining method. The optimal conditions for rice bran (68.1 g/l), peptone (9.1 g/l), and initial pH (7.0) of the medium for cell growth was determined by Design Expert Software based on the response surface method; conditions for production of CMCase were 55.2 g/l, 6.6 g/l, and 7.1, respectively. The optimal temperature for cell growth and the production of CMCase by B. atrophaeus LBH-18 was $30^{\circ}C$. The optimal conditions of agitation speed and aeration rate for cell growth in a 7-l bioreactor were 324 rpm and 0.9 vvm, respectively, whereas those for production of CMCase were 343 rpm and 0.6 vvm, respectively. The optimal inner pressure for cell growth and production of CMCase in a 100-l bioreactor was 0.06 MPa. Maximal production of CMCase under optimal conditions in a 100-l bioreactor was 127.5 U/ml, which was 1.32 times higher than that without an inner pressure. In this study, rice bran was developed as a carbon source for industrial scale production of CMCase by B. atrophaeus LBH-18. Reduced time for the production of CMCase from 7 to 10 days to 3 days by using a bacterial strain with submerged fermentation also resulted in increased productivity of CMCase and a decrease in its production cost.

• Korean Journal of Plant Taxonomy
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• v.47 no.3
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• pp.222-235
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• 2017

A comparative study of the leaf epidermal micromorphology in the tribe Neillieae (Neillia: 4 species, 4 varieties; Physocarpus: 5 species; Stephanandra: 2 species) was carried out using scanning electron microscopy in order to evaluate the taxonomic and systematic implications of these characteristics. The leaves of the genera Neillia and Stephanandra were hypostomatic, whereas those of P. monogynus, P. opulifolius were amphistomatic. The range of the size of the stomata is $12.02-34.39{\times}10.76-27.13{\mu}m$; the smallest was found in N. thyrsiflora (average $13.98{\times}12.43{\mu}m$; $L{\times}W$), while the largest was measured in N. gracilis (average $26.82{\times}20.67{\mu}m$; $L{\times}W$). Paracytic stomata complexes are only found in N. affinis, and the anomocytic type was most commonly found. The papillate epidermal cell type was only observed on the abaxial surfaces of P. insularis. Platelet epicuticular waxes were found on the adaxial surfaces of N. affinis and S. tanakae. Four types (unicellular non-glandular, two- to five-armed, stellate, and glandular) of trichomes were found on the leaves. Stellates were observed in all species of Physocarpus except for P. insularis. Consequently, leaf epidermal micromorphological characteristics (e.g., the presence of papillate epidermal cells and stellate, and stomata complexes) may have high taxonomic and systematic value in Neillieae. Our results strongly support previous molecular phylogenetic and palynological hypotheses that Stephanandra and Neillia are a single genus and that Physocarpus insularis should be considered as a member of Spiraea.

• Korean Journal of Breeding Science
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• v.42 no.5
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• pp.547-554
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• 2010

Low-molecular-weight glutenin subunit (LMW-GS) in common wheat (Triticum aestivum L.) is important for quality processing of bread and noodles. The objectives of this study were to clarify the composition of LMW-GSs and to identify their corresponding proteins. Using LMW-GS specific primers we cloned and characterized 43 LMW-GS genes in the wheat cultivar 'Jokyoung'. Some of these genes contain polypeptides different in size due to the presence of various deletions or insertions within repetitive and glutamine-rich domains. The comparison of deduced amino acid sequence of the LMW-GS genes in Jokyoung with that of 12 groups LMW-GSs of wheat cultivar Norin 61 showed that the deduced amino acid sequences were nearly the same to LMW-GS groups of 1, 2, 3/4, 5, 7, 10 and 11. All LMW-GS genes contain eight cysteine residues, which are conserved among all of the typical LMW-GS sequences. The relative positions of cysteine residues are also conserved, except those of the first and seventh. Based on phylogenetic analysis, the 43 sequences with the same N-terminal and C-terminal amino acid sequences were clustered in the same group. To identify the proteins containing the corresponding amino acid sequences, we determined the N-terminal amino acid sequence of 7 spots of LMW-GSs of Jokyoung separated by two-dimensional gel electrophoresis (2DE). Of them, Glu-B3 (LMW-m and LMW-s) and Glu-D3 (LMW-m) were detected in two and three spots, respectively and the others were not clear. Collectively, we classified diverse LMW-GSs and identified their corresponding protein products. These results will be helpful in breeding programs for improvement of wheat flour quality.

• Journal of Life Science
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• v.29 no.6
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• pp.679-687
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• 2019

Litsea populifolia, a plant species of the Lauraceae family, is widely distributed in the tropical and subtropical areas of Asia. The phylogenetic relationships and botanical characteristics of L. populifolia have been reported; however, its anti-oxidative and anti-cancer activities remain unclear. In this study, we evaluated the anti-oxidative and anti-cancer effects of ethanol extracts of L. populifolia (EELP) together with the molecular mechanism of its anti-cancer activity in human lung adenocarcinoma A549 cells. EELP showed significant anti-oxidative effects with a 50% inhibitory concentration at $11.71{\mu}g/ml$, which was measured by the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay. EELP exhibited cytotoxic activity and induced cell cycle arrest at the G1 phase in A549 cells in a dose-dependent manner, whereas EELP did not have the cytotoxic effect on the normal human lung cell line IMR90. Treatment with EELP also resulted in a decreased expression of G1/S transition-related molecules-including cyclin-dependent kinase (CDK) 2, CDK6, cyclin D1, and cyclin E-both for the transcription and translation levels. EELP-induced G1 arrest was associated with the phosphorylation of checkpoint kinase 2 (CHK2), p53, cell division cycle 25 homolog A (CDC25A), and the reduction of CDC25A expression in A549 cells. Collectively, these results suggest that EELP may exert an anti-cancer effect by cell cycle arrest at the G1 phase through both p53-dependent and p53-independent (ATM/CHK2/CDC25A/CDK2) pathways in A549 cells.

• Journal of Applied Biological Chemistry
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• v.62 no.3
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• pp.287-292
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• 2019

Pseudomonas tolaasii causes brown blotch disease on the oyster mushroom (Pleurotus ostreatus). Various pathogenic strains of P. tolaasii were isolated and divided into three subtypes, $P1{\alpha}$, $P1{\beta}$, and $P1{\gamma}$. For phage therapy, bacteriophages against to these subtype strains were applied to mushroom cultivation and very successful to prevent from the disease. In this study, bacteriophages were isolated against the representative strains of subtype pathogens and their polyclonal antibodies were synthesized to investigate structural relationship among capsid proteins of phages. Phage preparations over $10^{10}pfu/mL$ were injected to rabbit thigh muscle and polyclonal antibodies were obtained after three times of boost injection. Titers of the antibodies obtained were over $2{\times}10^7Ab/mL$ for the phage ${\phi}6264$, $1{\times}10^6Ab/mL$ for the phage ${\phi}HK2$, and $1{\times}10^7Ab/mL$ for the phage ${\phi}HK19$ and phage ${\phi}HK23$. High specific activities were observed between antibodies and the corresponding bacteriophages. Some cross-reactivities between the antibodies and non-corresponding bacteriophages were also measured. Antibody $Ab{\phi}6264$ inactivated all phages of $P1{\alpha}$ subtype and only phage ${\phi}HK16$ among $P1{\beta}$ subtype phages. Antibody $Ab{\phi}HK23$ of $P1{\gamma}$ subtype neutralized all phages of $P1{\beta}$ subtype as well as the phage ${\phi}HK23$, showing the widest phage-inactivation range. When the structural-similarity studies of phages were investigated by using phage antibodies, closeness obtained by phylogenetic analysis of 16S rRNA genes of pathogenic strains were quite different from that of polyclonal antibody-specific structural similarity of phage capsid proteins. In conclusion, there is weak correlation between the host strain specificity of bacteriophage and its capsid structural similarity measured by phage antibodies.

• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.552-558
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• 2021

Traditional foods are manufactured according to the characteristics of each region for the nations of the world. Korea has mainly farmed, and seasonings have developed around rice and vegetables. In fermented foods, lactic acid bacteria such as Lactobacillus sp. and Pediococcus sp. and Bacillus sp. were isolated and identified from fermented foods. In this study, lactic acid bacteria were isolated and identified from commercially available traditional Korean fermented foods, and candidate strains were selected through antibacterial activity tests on human and fish disease bacteria. Thereafter, the final strain was selected by examining the resistance to simulated gastric and intestinal fluids, and hemolysis. The three (M1, K1, C13) final selected latic acid bacteria were miced with prebiotics and the antibacterial activity of synbiotics was evaluated. As for the fist antibacterial activity result, C13 showed high antibacterial acitivity in human diseases and fish diseases. Then, M1, K1 and C13, which did not produce β-haemolysis and were resistant to simulated gastric and intestinal fluids, were subjected to the second antibacterial activity of synbiotics. When the three prebiotics (FOS, GOS, Inulin) and probiotics (M1, K1, C13) were mixed, the antibacterial activity was increased or inhibited. Based on the 16S rRNA gene sequencing results, K1 and M1 were analyzed as Bacillus tequiensis 99.72%, Bacillus subtilis 99.65%, Bacillus inaquosorum 99.72%, Bacillus cabrialesii 99.72%, Bacillus stercoris 99.58%, Bacillus spizizenii 99.58%, Bacillus halotolerans 99.58%, and Bacillus mojavensis 99.51%. And C13 was analyzed as Bacillus velezensis 99.71%, Bacillus nematocida 99.36%, Bacillus amyloliquefaciens 99.44%, Bacillus atrophaeus 99.22%, and Bacillus nakamurai 99.44%.

• Genomics & Informatics
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• v.21 no.3
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• pp.39.1-39.15
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• 2023

DNA barcoding without assessing reliability and validity causes taxonomic errors of species identification, which is responsible for disruptions of their conservation and aquaculture industry. Although DNA barcoding facilitates molecular identification and phylogenetic analysis of species, its availability in clariid catfish lineage remains uncertain. In this study, DNA barcoding was developed and validated for clariid catfish. 2,970 barcode sequences from mitochondrial cytochrome c oxidase I (COI) and cytochrome b (Cytb) genes and D-loop sequences were analyzed for 37 clariid catfish species. The highest intraspecific nearest neighbor distances were 85.47%, 98.03%, and 89.10% for COI, Cytb, and D-loop sequences, respectively. This suggests that the Cytb gene is the most appropriate for identifying clariid catfish and can serve as a standard region for DNA barcoding. A positive barcoding gap between interspecific and intraspecific sequence divergence was observed in the Cytb dataset but not in the COI and D-loop datasets. Intraspecific variation was typically less than 4.4%, whereas interspecific variation was generally more than 66.9%. However, a species complex was detected in walking catfish and significant intraspecific sequence divergence was observed in North African catfish. These findings suggest the need to focus on developing a DNA barcoding system for classifying clariid catfish properly and to validate its efficacy for a wider range of clariid catfish. With an enriched database of multiple sequences from a target species and its genus, species identification can be more accurate and biodiversity assessment of the species can be facilitated.

• Journal of Korean Society of Forest Science
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• v.86 no.3
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• pp.378-390
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• 1997

Ten total populations of Korean fir (Abies koreana Wilson) and Manshurian fir [A. nephrolepis (Traut.) Maxim.] were sampled from south Korea to investigate patterns of intraspecific variation in these species and to evaluate a recognition of the two species. Principal components analysis and cluster analysis were performed both on seed-cone data and on needle morphology data. The characters that contributed most to the separation between A. koreana and A. nephrolepis along three principal components axis were leaf width, length of seed, width of seed wing, length of seed wing, cone width, width of scale, and length of bract tip, but these characters were not diagnostic because of overlap in reality. Therefore, all these characters were not reliable in distinguishing these two taxa including bract position (exerted and recurved vs. exerted and straight). The individuals of A. koreana from Mt. Chi-ri appeared quite unique probably on account of its larger cone size and longer scale tip, while those from Mt. Hal-la of A. koreana were generally distinct from others in terms of their larger seed and seed wing and longer scale width. The Mt. Duk-yu specimens of A. korecana appeared somewhat smaller but more data were needed due to the small sampling size. Generally, the gradual clinal geographic trends made evident by the position of resin ducts in leaves of A. koreana can be detected. The southern populations, Mt. Hal-la (an insular population) were generally distinct from the northern populations (Mt. Chi-ri, Mt. Ga-ya and Mt. Duk-yu) in terms of their position of resin duct (medial, within mesophyll vs marginal, close to epidermis : 100% vs 75 or 50%). Although no sharp boundary separating these two species could be detected based on cone and needle morphology, the observed clinal pattern was distinct in northern populations of A. koreana and southern population of A. nephrnlepis. In a preceding study of the flavonoids variation of 20 species in eastern Asia, flavanone (5-deoxyflavanone) was found to be characteristic of A. faxoniana Rehder et Wilson, A. georgei Orr of China and A. koreana of Korea. A. faxoniana, which is assumed to be primitive species, has position of resin duct relative to both the medial and the marginal, while A. georgei and A. koreana are identified by marginal position of resin duct. With respect of foliar flavonoids chemistry, A. koreana was distinct from A. nephrolepis : the southmost samples (Mt. Hal-la and Mt. Chi-ri) contained additional flavonoids derivatives (mainly flavanone) that were not found in the northmost samples of A. nephrolepis except a few individuals from Mts. Seo-rak and Tae-bak populations of Kwang-won province. The presence of A. koreana type flavonoids in two Chinese species suggested that position of resin duct may be a phyletic character. Abies koreana including two Chinese taxa, exhibited the most elaborate and specialized flavonoids profile within the Abies in eastern Asia. Contrary to our initial expectations, the apparent intermediates between A. nephrolepis and A. koreana in Duk-yu and Ga-ya mountains were found. The pattern of variation on position of resin duct and flavonoids chemistry in these populations of A. kareana suggested that genetic interchange or natural hybridization had occurred between these two species. The evidence needed to resolve the status of this taxon is still inconclusive in our opinion until intermediate individuals from Mts. Duk-yu and Ga-ya show indication of hybridization between the two species.