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Cloning of α-Amylase Gene from Unculturable Bacterium Using Cow Rumen Metagenome (소 반추위 메타게놈에서 비배양 세균의 α-amylase 유전자 클로닝)

  • Cho, Soo-Jeong;Yun-Han-Dae
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.1013-1021
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    • 2005
  • The metagenomes of complex microbial communities are rich sources of novel biocatalysts. The gene encoding an extracellular $\alpha$-amylase from a genomic DNA of cow rumen was cloned in Escherichia coli DH5$\alpha$ and sequenced. The $\alpha$-amylase (amyA) gene was 1,893 bp in length, encoding a protein of 631 amino acid residues with calculated molecular weight of 70,734 Da. The molecular weight of the enzyme was estimated to be about 71,000 Da by active staining of a SDS-PACE. The enzyme was 21 to $59\%$ sequence identical with other amyloyltic enzymes. The AmyA was optimally active at pH 6.0 and $40\%$. The AmyA had a calculated pI of 5.87. AmyA expressed in E. coli DH5$\alpha$ was enhanced in the presence of $Mg^{2+}$ (20 mM) and $Ca^{2+}$ (30 mM) and inhibited in the presence of $Fe^{2+}$ and $Cu^{2+}$. The origin of amyA gene could not be confirmed by PCR using internal primer of amyA gene from extracted genomic DNA of 49 species rumen culturable bacteria so far. An amyh is supposed to obtained from unculturable rumen bacterium in cow rumen environment.

The Improvement of Fabrication Process for a-Si:H TFT's Yield (a-Si:H TFT의 수율 향상을 위한 공정 개선)

  • Hur, Chang-Wu
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.11 no.6
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    • pp.1099-1103
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    • 2007
  • TFT's have been intensively researched for possible electronic and display applications. Through tremendous engineering and scientific efforts, a-Si:H TFT fabrication process was greatly improved. In this paper, the reason on defects occurring at a-Si:H TFT fabrication process is analyzed and solved, so a-Si:H TFT's yield is increased and reliability is improved. The a-Si:H TFT of this paper is inverted staggered type TFT. The gate electrode is formed by patterning with length of $8{\mu}m{\sim}16{\mu}m$ and width of $80{\sim}200{\mu}m$ after depositing with gate electrode (Cr). We have fabricated a-SiN:H, conductor, etch-stopper and photo-resistor on gate electrode in sequence, respectively. We have deposited n+a-Si:H, NPR(Negative Photo Resister) layer after forming pattern of Cr gate electrode by etch-slower pattern. The NPR layer by inverting pattern of upper Sate electrode is patterned and the n+a-Si:H layer is etched by the NPR pattern. The NPR layer is removed. After Cr layer is deposited and patterned, the source-drain electrode is formed. The a-Si:H TFT made like this has problems at photo-lithography process caused by remains of PR. When sample is cleaned, this remains of PR makes thin chemical film on surface and damages device. Therefor, in order to improve this problem we added ashing process and cleaning process was enforced strictly. We can estimate that this method stabilizes fabrication process and makes to increase a-Si:H TFT's yield.

Five Newly Collected Turnip Mosaic Virus (TuMV) Isolates from Jeju Island, Korea are Closely Related to Previously Reported Korean TuMV Isolates but Show Distinctive Symptom Development

  • Hu, Wen-Xing;Kim, Byoung-Jo;Kwak, Younghwan;Seo, Eun-Young;Kim, Jung-Kyu;Han, Jae-Yeong;Kim, Ik-Hyun;Lim, Yong Pyo;Cho, In-Sook;Domier, Leslie L;Hammond, John;Lim, Hyoun-Sub
    • The Plant Pathology Journal
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    • v.35 no.4
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    • pp.381-388
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    • 2019
  • For several years, temperatures in the Korean peninsula have gradually increased due to climate change, resulting in a changing environment for growth of crops and vegetables. An associated consequence is that emerging species of insect vector have caused increased viral transmission. In Jeju Island, Korea, occurrences of viral disease have increased. Here, we report characterization of five newly collected turnip mosaic virus (TuMV) isolates named KBJ1, KBJ2, KBJ3, KBJ4 and KBJ5 from a survey on Jeju Island in 2017. Full-length cDNAs of each isolate were cloned into the pJY vector downstream of cauliflower mosaic virus 35S and bacteriophage T7 RNA polymerase promoters. Their fulllength sequences share 98.9-99.9% nucleotide sequence identity and were most closely related to previously reported Korean TuMV isolates. All isolates belonged to the BR group and infected both Chinese cabbage and radish. Four isolates induced very mild symptoms in Nicotiana benthamiana but KBJ5 induced a hypersensitive response. Symptom differences may result from three amino acid differences uniquely present in KBJ5; Gly(382)Asp, Ile(891)Val, and Lys(2522)Glu in P1, P3, and NIb, respectively.

Molecular Phylogeny of Chattonella (Raphidophyceae) Species from Deungnyang Bay, Korea Using Single-Cell PCR (Single-cell PCR을 이용하여 분석한 득량만 Chattonella 종 (Raphidophyceae)의 분자계통학적 특성)

  • Kim, Jin Joo;Song, Seon Yeung;Park, Tae Gyu
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.24 no.7
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    • pp.967-972
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    • 2018
  • The genus Chattonella belonging to the class raphidophyceae, is a harmful algal bloom species. Recently, its occurrence has been increasing and expanding along the Korean coast. Species identification of the genus Chattonella only by morphological observation is difficult due to the lack of rigid cell walls. In this study, the morphological characteristics and genetic affinity of Chattonella sp. isolated from Deungnyang Bay in 2017 were examined. We carried out single-cell isolation from field samples then sequenced three different areas using the single-cell PCR method: 1) parts of ribosomal operon, the large subunit (LSU) of the rDNA, 2) the chloroplast-encoded subunit psaA of Photosystem I, and 3) rbcL encoding the large subunit of the Rubisco gene. The cells were morphologically very similar to the general genus Chattonella ($74.0{\pm}10.1{\mu}m$ in length, $33.1{\pm}3.6{\mu}m$ in width). The three partial gene sequences were insufficient to justify distinction at the species rank. However, they clustered at 99-100 % sequence similarity with C. marina, C. marina var. antiqua and C. marina var. ovata.

Study of the method of production of excavated arrow bundle and its conservation treatment (발굴 출토 화살다발 제작기법 연구 및 보존처리)

  • Lee, Byeonghoon;Choi, Bobae;Huh, Ilgwon
    • Conservation Science in Museum
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    • v.25
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    • pp.9-26
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    • 2021
  • This paper describes the production methods that were originally used for an arrow bundle excavated from a Bronze Age residential area in Auraji in Jeongseon, Gangwon-do Province and the conservation treatment process that it subsequently underwent. An arrow conventionally consists of an arrowhead and a shaft. It is rare to excavate a shaft along with an arrowhead in a complete form since the shaft is made of organic materials. Notably, the arrow bundle from the Auraji site is of great significance as it shows traces of tangless stone arrowheads attached to charred shafts and offers an important case of the split end of a piece of a tree being inserted into an arrowhead. For a further examination of the characteristics of the arrows from Auraji, microscopic investigation was conducted and the type of wood used for the arrow shafts was examined. The sequence and direction of processing and the particle sizes of the grinding tools were revealed through the analysis of traces of grinding on the stone arrowheads. The shaft is presumed to have been made from a green length of three-year-old willow (Salix spp.). A curing agent with a high degree of waterproofing and reversibility was used during the on-site curing process according to demands of the surrounding environment, and a technique that the authors call the "Bridge" method was used for emergency collection of the relics. Once the bundle was transferred to the conservation treatment lab, reinforcing materials were carefully chosen as it was important not to damage the relics during the process of turning them for the repair of their reverse sides. For this purpose, artificial clay was selected since it can safely bear a load and has excellent physical properties. Finally, detached parts were rejoined, the relics and their surrounding materials were cleaned, and the bottom sides were finished with epoxy resin prior to the display of the relics at the museum.

Characterization of the Novel Marine Bacterium Planococcus sp. 107-1T (신종 해양미생물 Planococcus sp. 107-1T의 분류학적 특성 분석)

  • Kim, Dong-Gyun;Jung, Hyun-Kyoung;Kim, Young-Ok;Kong, Hee Jeong;Nam, Bo-Hye;Kim, Ju-Won;Kim, Young-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.55 no.5
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    • pp.612-624
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    • 2022
  • A novel Gram-positive, motile, non-spore forming aerobic marine bacterium, designated 107-1T was isolated from tidal mud collected in Gyehwa-do, South Korea. Cells of strain 107-1T were short rod or coccoid, oxidase negative, catalase positive and grew at 10-40℃ (with optimum growth at 25-30℃). It utilized menaquinones MK-7 and 8 as its respiratory quinones and its major fatty acids were anteiso-C15:0 (37.9%), iso-C16:0 (14.9%), and iso-C14:0 (10.8%). Phylogenetic analysis based on 16S rRNA gene sequences revealed a distinct clade containing strain 107-1T and close species Planococcus ruber CW1T(98.52% sequence similarity), P. faecalis KCTC 33580T(97.67%), P. kocurii ATCC 43650T(97.48%), P. donghaensis DSM 22276T(97.47%), and P. halocryophilus DSM 24743T(97.37%). Strain 107-1T contains one circular chromosome (3,513,248bp in length) with G+C content of 44.6 mol%. Estimated ranges for genome to genome distance, average nucleotide identity, and average amino acid identity comparing strain 107-1T with close taxa were 20.3-34.8%, 77.9-86.9%, and 73.6-92.8%, respectively. Based on polyphasic analysis, strain 107-1T represents a novel species belonging to the genus Planococcus.

Control of the flow past a sphere in a turbulent boundary layer using O-ring

  • Okbaz, Abdulkerim;Ozgoren, Muammer;Canpolat, Cetin;Sahin, Besir;Akilli, Huseyin
    • Wind and Structures
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    • v.35 no.1
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    • pp.1-20
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    • 2022
  • This research work presents an experimental study's outcomes to reveal the impact of an O-ring on the flow control over a sphere placed in a turbulent boundary layer. The investigation is performed quantitatively and qualitatively using particle image velocimetry (PIV) and dye visualization. The sphere model having a diamater of 42.5 mm is located in a turbulent boundary layer flow over a smooth plate for gap ratios of 0≤G/D≤1.5 at Reynolds number of 5 × 103. Flow characteristics, including patterns of instantaneous vorticity, streaklines, time-averaged streamlines, velocity vectors, velocity fluctuations, Reynolds stress correlations, and turbulence kinetic energy (), are compared and discussed for a naked sphere and spheres having O-rings. The boundary layer velocity gradient and proximity of the sphere to the flat plate profoundly influence the flow dynamics. At proximity ratios of G/D=0.1 and 0.25, a wall jet is formed between lower side of the sphere and flat plate, and velocity fluctuations increase in regions close to the wall. At G/D=0.25, the jet flow also induces local flow separations on the flat plate. At higher proximity ratios, the velocity gradient of the boundary layer causes asymmetries in the mean flow characteristics and turbulence values in the wake region. It is observed that the O-ring with various placement angles (𝜃) on the sphere has a considerable alteration in the flow structure and turbulence statistics on the wake. At lower placement angles, where the O-ring is closer to the forward stagnation point of the sphere, the flow control performance of the O-ring is limited; however, its impact on the flow separation becomes pronounced as it is moved away from the forward stagnation point. At G/D=1.50 for O-ring diameters of 4.7 (2 mm) and 7 (3 mm) percent of the sphere diameter, the -ring exhibits remarkable flow control at 𝜃=50° and 𝜃=55° before laminar flow separation occurrence on the sphere surface, respectively. This conclusion is yielded from narrowed wakes and reductions in turbulence statistics compared to the naked sphere model. The O-ring with a diameter of 3 mm and placement angle of 50° exhibits the most effective flow control. It decreases, in sequence, streamwise velocity fluctuations and length of wake recovery region by 45% and 40%, respectively, which can be evaluated as source of decrement in drag force.

Development and Verification of Smart Greenhouse Internal Temperature Prediction Model Using Machine Learning Algorithm (기계학습 알고리즘을 이용한 스마트 온실 내부온도 예측 모델 개발 및 검증)

  • Oh, Kwang Cheol;Kim, Seok Jun;Park, Sun Yong;Lee, Chung Geon;Cho, La Hoon;Jeon, Young Kwang;Kim, Dae Hyun
    • Journal of Bio-Environment Control
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    • v.31 no.3
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    • pp.152-162
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    • 2022
  • This study developed simulation model for predicting the greenhouse interior environment using artificial intelligence machine learning techniques. Various methods have been studied to predict the internal environment of the greenhouse system. But the traditional simulation analysis method has a problem of low precision due to extraneous variables. In order to solve this problem, we developed a model for predicting the temperature inside the greenhouse using machine learning. Machine learning models are developed through data collection, characteristic analysis, and learning, and the accuracy of the model varies greatly depending on parameters and learning methods. Therefore, an optimal model derivation method according to data characteristics is required. As a result of the model development, the model accuracy increased as the parameters of the hidden unit increased. Optimal model was derived from the GRU algorithm and hidden unit 6 (r2 = 0.9848 and RMSE = 0.5857℃). Through this study, it was confirmed that it is possible to develop a predictive model for the temperature inside the greenhouse using data outside the greenhouse. In addition, it was confirmed that application and comparative analysis were necessary for various greenhouse data. It is necessary that research for development environmental control system by improving the developed model to the forecasting stage.

Chloroplast genome sequence and PCR-based markers for S. cardiophyllum (감자 근연야생종 Solanum cardiophyllum의 엽록체 전장유전체 구명 및 이를 이용한 S. cardiophyllum 특이적 분자마커의 개발)

  • Tae-Ho Park
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.45-55
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    • 2023
  • The diploid Solanum cardiophyllum, a wild tuberbearing species from Mexico is one of the relatives to potato, S. tuberosum. It has been identified as a source of resistance to crucial pathogens and insects such as Phytophthora infestans, Potato virus Y, Colorado potato beetle, etc. and is widely used for potato breeding. However, the sexual hybridization between S. cardiophyllum and S. tuberosum is limited due to their incompatibility. Therefore, somatic hybridization can introduce beneficial traits from this wild species into the potato. After somatic hybridization, selecting fusion products using molecular markers is essential. In the current study, the chloroplast genome of S. cardiophyllum was sequenced by next-generation sequencing technology and compared with those of other Solanum species to develop S. cardiophyllum-specific markers. The total length of the S. cardiophyllum chloroplast genome was 155,570 bp and its size, gene content, order and orientation were similar to those of the other Solanum species. Phylogenic analysis with 32 other Solanaceae species revealed that S. cardiophyllum was expectedly grouped with other Solanum species and most closely located with S. bulbocastanum. Through detailed comparisons of the chloroplast genome sequences of eight Solanum species, we identified 13 SNPs specific to S. cardiophyllum. Further, four SNP-specific PCR markers were developed for discriminating S. cardiophyllum from other Solanum species. The results obtained in this study would help to explore the evolutionary aspects of Solanum species and accelerate breeding using S. cardiophyllum.

A Taxonomic Reconsideration of the Genus Lemna L. (Lemnaceae) in Korea (한국산 좀개구리밥속(개구리밥과)의 분류학적 실체에 대한 재고)

  • Kim, Yong-In;Shim, Sang In;Park, Jin Hee
    • Korean Journal of Environment and Ecology
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    • v.31 no.4
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    • pp.349-364
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    • 2017
  • Duckweed family (Lemnaceae Martinov), including the genus Lemna L., is a typical floating aquatic perennial plant, and about five genera and 40 species in the family are in wide distribution around the world except the polar regions. The genus Lemna is the smallest and the simplest plant among the angiosperms. It has a characteristic of doubling every three days with fast vegetative propagation, which helps the organisms to increase in rapid growth. As such, the plant is ideal for environmental pollution assessment and toxicity test. Although taxonomists and scholars have used different scientific names for the species, many of them have agreed that there is only one member of species of the genus Lemna in Korea. Paying attention to the external morphological variation observed in the Korean genus Lemna, we conducted a molecular phylogenetic analysis to identify the entity of the Korean Lemna species and to investigate the possibility of two or more members of the species existing in Korea. We determined and aligned the DNA sequences of the atpF-H region of the chloroplast DNA in 37 populations of the nationally distributed Lemna species. The results showed that the sequence length of the cp DNA atpF-H region was 463-483 bp, the length of the aligned sequences was 488 bp, and the number of variation site in nucleotide sequences was 47. There were two types of aligned sequences of the cp DNA atpF-H region from 37 populations of Lemna species in Korea. The maximum parsimony analysis revealed that the Korean Lemna consists of two clades, and one of them had two subclades. The results suggest that, contrary to the general understanding, at least two taxa (L.aequinoctialis, L.minor) exist in Korea.