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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.

Antioxidant Effects of Cysteine-containing Peptides of Different Lengths in Human HaCaT Keratinocytes Exposed to Hydrogen Peroxide (과산화수소에 노출된 인간 각질형성세포에서 길이가 다른 시스테인 함유 펩타이드의 항산화 효과)

  • Jae Won Ha;Joon Yong Choi;Yong Chool Boo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.3
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    • pp.193-201
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    • 2023
  • Hydrogen peroxide (H2O2) is a type of active oxygen species (ROS) that causes oxidative stress in cells and affects cell growth, proliferation, senescence, and death. The purpose of this study is to find active peptides that attenuate cytotoxicity of H2O2. A positional scanning synthetic tetrapeptide combinatorial library was screened to predict the sequence of potentially active peptides. As a result of comparing the effect of peptide pools on H2O2-induced death of human keratinocytes (HaCaT cells), various active peptide sequences were predicted. Especially, peptides containing cysteine (C) residue were predicted to be active. In follow-up experiments, the cytotoxicity and activity of cysteine-containing peptides of different lengths, such as C-NH2, CC-NH2, CCC-NH2, and CCCC-NH2 were examined. C-NH2 and CC-NH2 showed no significant cytotoxicity up to 1.0 mM, but CCC-NH2, and CCCC-NH2 showed relatively strong cytotoxicity. C-NH2 and CC-NH2 alleviated H2O2-induced cytotoxicity. CC-NH2 was more cytoprotective compared to C-NH2, C, N-acetyl cysteine (NAC), and glutathione (GSH). When intracellular ROS was measured by flow cytometry, H2O2 increased ROS production, and CC-NH2 suppressed ROS production more effectively than C-NH2, and it was as effective as C, NAC, and GSH. This study suggests that CC-NH2 of the cysteine-containing peptides of different lengths has an antioxidant property that safely and effectively alleviates H2O2-induced cytotoxicity and ROS production.

Efficiency of Density Gradient Centrifugation Method (Ludox method) Based on eDNA for the Analysis of Harmful Algal Bloom Potential (유해남조류 발생 잠재성 분석을 위한 eDNA 기반의 퇴적물 전처리 방법: 밀도 구배 원심분리법(Ludox method))

  • Kyeong-Eun Yoo;Hye-In Ho;Hyunjin Kim;Keonhee Kim;Soon-Jin Hwang
    • Korean Journal of Ecology and Environment
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    • v.56 no.1
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    • pp.36-44
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    • 2023
  • Environmental DNA (eDNA) can exist in both intracellular and extracellular forms in natural ecosystems. When targeting harmful cyanobacteria, extracellular eDNA indicates the presence of traces of cyanobacteria, while intracellular eDNA indicates the potential for cyanobacteria to occur. However, identifying the "actual" potential for harmful cyanobacteria to occur is difficult using the existing sediment eDNA analysis method, which uses silica beads and cannot distinguish between these two forms of eDNA. This study analyzes the applicability of a density gradient centrifugation method (Ludox method) that can selectively analyze intracellular eDNA in sediment to overcome the limitations of conventional sediment eDNA analysis. PCR was used to amplify the extracted eDNA based on the two different methods, and the relative amount of gene amplification was compared using electrophoresis and Image J application. While the conventional bead beating method uses sediment as it is to extract eDNA, it is unknown whether the mic gene amplified from eDNA exists in the cyanobacterial cell or only outside of the cell. However, since the Ludox method concentrates the intracellular eDNA of the sediment through filtration and density gradient, only the mic gene present in the cyanobacteria cells could be amplified. Furthermore, the bead beating method can analyze up to 1 g of sediment at a time, whereas the Ludox method can analyze 5 g to 30 g at a time. This gram of sediments makes it possible to search for even a small amount of mic gene that cannot be searched by conventional bead beating method. In this study, the Ludox method secured sufficient intracellular gene concentration and clearly distinguished intracellular and extracellular eDNA, enabling more accurate and detailed potential analysis. By using the Ludox method for environmental RNA expression and next-generation sequencing (NGS) of harmful cyanobacteria in the sediment, it will be possible to analyze the potential more realistically.

Feasibility of Environmental DNA Metabarcoding for Invasive Species Detection According to Taxa (분류군별 외래생물 탐지를 위한 환경 DNA 메타바코딩 활용 가능성)

  • Yujin Kang;Jeongeun Jeon;Seungwoo Han;Suyeon Won;Youngkeun Song
    • Journal of Environmental Impact Assessment
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    • v.32 no.2
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    • pp.94-111
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    • 2023
  • In order to establish an effective management strategy for invasive species early detection and regular monitoring are required to assess their introduction or dispersal. Environmental DNA (eDNA) is actively applied to evaluate the fauna including the presence of invasive species as it has high detection sensitivity and can detect multiple species simultaneously. In Korea, the applicability evaluation of metabarcoding is being conducted mainly on fish, and research on other taxa is insufficient. Therefore, this study identified the feasibility of detecting invasive species in Korea using eDNA metabarcoding. In addition, to confirm the possibility of detection by taxa, the detection of target species was evaluated using four universal primers (MiFish, MiMammal, Mibird, Amp16S) designed for fish, mammals, birds, and amphibians. As a result, target species (Trachemys scripta, 3 sites; Cervus nippon, 3 sites; Micropterus salmoides, 7 sites; Rana catesbeiana, 4 sites) were detected in 17 of the total 55 sites. Even in the selection of dense sampling sites within the study area, there was a difference in the detection result by reflecting the ecological characteristics of the target species. A comparison of community structures (species richness, abundance and diversity) based on the presence of invasive species focused on M.salmoides and T.scripta, showed higher diversity at the point where invasive species were detected. Also, 1 to 4 more species were detected and abundance was also up to 1.7 times higher. The results of invasive species detection through metabarcoding and the comparison of community structures indicate that the accumulation of large amounts of monitoring data through eDNA can be efficiently utilized for multidimensional ecosystem evaluation. In addition, it suggested that eDNA can be used as major data for evaluation and prediction, such as tracking biological changes caused by artificial and natural factors and environmental impact assessment.

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

  • Jin Sun Kim;Dong-Geun Lee;Go-Wun Yeo;Min-Joo Park;Sang-Hyeon Lee
    • Journal of Life Science
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    • v.33 no.4
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    • pp.357-362
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    • 2023
  • This report looks at an agar-degrading marine bacterium and characterization of its agarase. Agar-degrading marine bacterium JS-1 was isolated with Marine agar 2216 media from seawater from the seashore of Sojuk-do, Changwon in Gyeongnam Province, Korea. The agar-degrading bacterium was named as Agarivorans sp. JS-1 by phylogenetic analysis based on 16S rRNA gene sequencing. The extracellular agarase was prepared from the culture media of Agarivorans sp. JS-1 and used for characterization. Relative activities at 20℃, 30℃, 35℃, 40℃, 45℃, 50℃, 55℃, and 60℃ were 70%, 74%, 78%, 83%, 87%, 100%, 74%, and 66%, respectively. Relative activities at pH 5, 6, 7, and 8 were 91%, 100%, 90%, and 89%, respectively. Its extracellular agarase showed maximum activity (207 units/l) at pH 6.0 and 50℃ in 20 mM Tris-HCl buffer. The residual activity after heat treatment at 20℃, 30℃, and 50℃ for 30 minutes was 90%, 70%, and 50% or more, respectively. After a 2-hour heat treatment at 20℃, 30℃, 35℃, 40℃, and 45℃, the residual activity was 80%, 68%, 65%, 63%, and 57%, respectively. At 50℃ and above, after heat treatment for 30 minutes, the residual activity was below 60%. Thin layer chromatography analysis suggested that Agarivorans sp. JS-1 produces extracellular β-agarases as they hydrolyze agarose to produce neoagarooligosaccharides such as neoagarohexaose (20.6%), neoagarotetraose (58.5%), and neoagarobiose (20.9%). Agarivorans sp. JS-1 and its thermotolerant β-agarase would be useful in the production of neoagarooligosaccharides, showing functional activity such as inhibition of bacterial growth and delay of starch degradation.

Isolation and Characterization of the Indigenous Microalgae Chlamydomonas reinhardtii K01 as a Potential Resource for Lipid Production and Genetic Modification (지질생산 및 유전자 조작의 잠재적 자원으로서의 토착 미세조류 Chlamydomonas reinhardtii K01의 분리 및 특성)

  • Kim, Eun-Kyung;Cho, Dae Hyun;Suh, Sang-Ik;Lee, Chang-Jun;Kim, Hee-Sik;Suh, Hyun-Hyo
    • Journal of Life Science
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    • v.32 no.3
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    • pp.202-209
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    • 2022
  • The green alga Chlamydomonas reinhardtii, a unicellular haploid eukaryote, has long been used by researchers and industries as a cell factory to produce high value-added microalgae substances using genetic modification. Microalga K01, presumed to be Chlamydomonas, was isolated from 12 freshwater samples from the Chungcheong and Jeolla regions to replace C. reinhardtii, an introduced species currently used in most basic and industrial research. The isolated K01 strain was identified as C. reinhardtii through morphological and phylogenetic studies of the 18S rDNA gene sequence (NCBI accession number KC166137). The growth and lipid content of the isolated C. reinhardtii K01 were compared with three wild and four mutant strains in TAP medium, and it was found that the K01 strain could produce 1.74×107 cells/ml by the third day of culture. The growth rate of C. reinhardtii K01 was 1.5 times faster than UTEX2244, which showed the highest number of cells (1.20×107 cells/ml) among the compared strains. The lipid content of the isolated C. reinhardtii K01 (20.67%) was similar to those of the wild strains, although the fatty acid oleate C18:1 was not detected in the isolated strain but was identified in the seven others. The cell density of the isolated strain increased to 0.87 g/l during a six-day culture in BG11 medium, where nitrate (NaNO3) was introduced as a nitrogen source, while the seven acquired strains showed almost no cell proliferation.

Fecal Microbiota Profiling of Holstein and Jersey, in South Korea : A Comparative Study (국내에서 사육되는 Holstein 젖소과 Jersey 젖소의 대변 미생물 분석 : 비교연구)

  • Gwangsu Ha;Ji-Won Seo;Hee Gun Yang;Se Won Park;Soo-Young Lee;Young Kyoung Park;RanHee Lee;Do-Youn Jeong;Hee-Jong Yang
    • Journal of Life Science
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    • v.33 no.7
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    • pp.565-573
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    • 2023
  • In light of the complex interactions between the host animal and its resident gut microbiomes, studies of these microbial communities as a means to improve cattle production are important. This study was conducted to analyze the intestinal microorganisms of Holstein (HT) and Jersey (JS), raised in Korea and to clarify the differences in microbial structures according to cattle species through next-generation sequencing. The alpha-diversity analysis revealed that most species richness and diversity indices were significantly higher in JS than in HT whereas phylogenetic diversity, which is the sum of taxonomic distances, is not significant. Microbial composition analysis showed that the intestinal microbial community structure of the two groups differed. In the both groups, a significant correlation was observed among the distribution of several microbes at the family level. In particular, a highly significant correlation (p<0.0001) among a variety of microbial distributions was found in JS. Beta-diversity analyis was to performed to statistically verify whether a difference exists in the intestinal microbial community structure of the two groups. Principal coordinate analysis and unweighted pair group method with arithmetic mean (UPGMA) clustering analysis showed separation between the HT and JS clusters. Meanwhile, permutational multivariate analysis of variance (PERMANOVA) revealed that their microbial structures are significantly different (p<0.0001). LEfSe biomarker analysis was performed to discover the differenc microbial features between the two groups. We found that several microbes, such as Firmicutes, Bacilli, Moraxellaceae and Pseudomonadales account for most of the difference in intestinal microbial community structure between the two groups.

Development of Genetic Selection Marker via Examination of Genome in Bacillus velezensis K10 (Bacillus velezensis K10 유전체 분석을 통한 균주 선발 마커 개발)

  • Sam Woong Kim;Young Jin Kim;Tae Wook Lee;Won-Jae Chi;Woo Young Bang;Tae Wan Kim;Kyu Ho Bang;Sang Wan Gal
    • Journal of Life Science
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    • v.33 no.11
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    • pp.897-904
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    • 2023
  • This study was done to develope genetic markers with the unique characteristics of genes according to the genomic information of Bacillus velezensis K10. B. velezensis K10 maintained a total of 4,159,835 bps, which was found to encode 5,136 open reading frames (orfs). B. velezensis K10 was found to have much more gene migration due to external factors overall compared to standard strain B. velezensis JS25R. In order to discover genetic selection markers, orfs on the genome to be easily induced to gene mutation were surveyed such as recombinase, integrase, transposase, and phage-related genes. As a result of the investigation, 9 candidate markers were isolated with high possibility as genetic selection markers. Although a part in the various origin's areas showed specificities in comparison with homology, the selected markers were all existed in phage-related areas because they were relatively lower homologies in phage-related genes. PCR analysis was done on B. licheniformis K12, B. velezensis K10, B. subtilis, and B. cereus to establish them as inter-species candidate selection markers. As a result, it was confirmed that B. velezensis K10-specific PCR products were formed in a total of 6 primer sets such as BV3 and BV5 to 9. On the other hand, analysis at the subspecies level observed the formation of B. velezensis K10-specific PCR products in 4 primer sets such as BV3, 5, 8, and 9. Among them, since BV5 and BV8 were detected by very specific results, we suggest that BV5 and 8 can be used as B. velezensis K10 gene selection markers at the species and sub-species level.

A Study on the Revitalization of the Competency Assessment System in the Public Sector : Compare with Private Sector Operations (공공부문 역량평가제도의 활성화 방안에 대한 연구 : 민간부분의 운영방식과의 비교 연구)

  • Kwon, Yong-man;Jeong, Jang-ho
    • Journal of Venture Innovation
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    • v.4 no.1
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    • pp.51-65
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    • 2021
  • The HR policy in the public sector was closed and operated mainly on written tests, but in 2006, a new evaluation, promotion and education system based on competence was introduced in the promotion and selection system of civil servants. In particular, the seniority-oriented promotion system was evaluated based on competence by operating an Assessment Center related to promotion. Competency evaluation is known to be the most reliable and valid evaluation method among the evaluation methods used to date and is also known to have high predictive feasibility for performance. In 2001, 19 government standard competency models were designed. In 2006, the competency assessment was implemented with the implementation of the high-ranking civil service team system. In the public sector, the purpose of the competency evaluation is mainly to select third-grade civil servants, assign fourth-grade civil servants, and promotion fifth-grade civil servants. However, competency assessments in the public sector differ in terms of competency assessment objectives, assessment processes and competency assessment programmes compared to those in the private sector. For the purposes of competency assessment, the public sector is for the promotion of candidates, and the private sector focuses on career development and fostering. Therefore, it is not continuously developing capabilities than the private sector and is not used to enhance performance in performing its duties. In relation to evaluation items, the public sector generally operates a system that passes capacity assessment at 2.5 out of 5 for 6 competencies, lacks feedback on what competencies are lacking, and the private sector uses each individual's competency score. Regarding the selection and operation of evaluators, the public sector focuses on fairness in evaluation, and the private sector focuses on usability, which is inconsistent with the aspect of developing capabilities and utilizing human resources in the right place. Therefore, the public sector should also improve measures to identify outstanding people and motivate them through capacity evaluation and change the operation of the capacity evaluation system so that they can grow into better managers through accurate reports and individual feedback

Genetic Characterization of Antigenic Variant Infectious Bursal Disease Virus (IBDV) in Chickens in Korea

  • Jong-Yeol Park;Ki-Woong Kim;Ke Shang;Sang-Won Kim;Yu-Ri Choi;Cheng-Dong Yu;Ji-Eun Son;Gyeong-Jun Kim;Won-Bin Jeon;In-Hwan Kim;Bai Wei;Min Kang;Hyung-Kwan Jang;Se-Yeoun Cha
    • Korean Journal of Poultry Science
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    • v.50 no.4
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    • pp.231-240
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    • 2023
  • Infectious bursal disease (IBD) is an acute, highly contagious, and immunosuppressive disease in young chickens, and causes considerable economic losses to the poultry industry. More than 30 years ago, an antigenic variant IBDV (avIBDV) was reported in chicken farms in the United States. Recently, a novel avIBDV exhibited clear differences in molecular characteristics compared with previous variant strains. This study investigated the molecular characteristics of recently isolated avIBDV strains in Korea. Strains of avIBDV were confirmed by reverse transcription PCR (RT-PCR) and were propagated in 10-day-old specific-pathogen-free (SPF) embryonated chicken eggs through chorioallantoic membrane (CAM) inoculation. Multiple sequence alignment and phylogenetic analyses of hypervariable regions VP2 gene revealed that the strains originated from two different avIBDV lineages (G2a and G2d). In our results, we confirmed the co-existence and prevalence of avIBDV genogroup G2a and G2d in chicken farms. It is necessary to study the protective efficacy of current vaccines against avIBDVs.