• Title/Summary/Keyword: RNA 1 structure

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Molecular Cloning and Identification of Novel Genes, Gomsin, Characteristically Expressed in Snailfish, Liparis tanakae (꼼치에서 특징적으로 발현되는 새로운 유전자 곰신의 분리 및 동정)

  • 송인선;이석근;손진기
    • Development and Reproduction
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    • v.6 no.1
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    • pp.7-16
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    • 2002
  • In order to obtain the specific genes of snailfish a subtracted cDNA library was constructed, and analysed by sequencing and GenBank search. Among them C90-171 clone was turned out to be genes showing low homology and nonredundant genes. This novel clone was named Gomsin(C90-171). Gomsin was shown to be intensely expressed in the epithelial cells, some mesenchymal cells, and sheaths of muscle bundles in the result of immunohistochemistry. In the cross reaction assay of Gomsin antibody against various human tissues, the Gomsin was strongly expressed in the ductal and acinar cells of salivary glands, which was similar to the expression patterns of proline-rich proteins(PRPs) of human. The antibody raised against the Gomsin was clearly cross-reacted with human salivary PRPs and also recombinant proteins of human PRPs in the Western blot and immunoprecipitation analysis. Contrast to the salivary PRPs, the Gomsin was not easily degraded in the mixed saliva, but rapidly attacked on the cultured keratocytes in vitro. The simulated protein structure of Gomsin was similar to the whorled pattern of PRPs, even though the amino acid sequence of Gomsin was quite different from those of PRPs. These data suggest that the Gomsin is a characteristic matrix protein in the skin and body of snailfish, which is also utilized for the tissue protection in the similar way to the PRPs of human muco-secretory organs.

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High-rate Denitrifying Process Based on Methanol and Characteristics of Organic Carbon Uptake (메탄올 기반 탈질 공정의 고속화 및 탄소 섭취 특성)

  • Park, Suin;Jeon, Junbeom;Bae, Hyokwan
    • Journal of Korean Society on Water Environment
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    • v.36 no.6
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    • pp.581-591
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    • 2020
  • In this study, two types of reactors were operated to examine the properties of methanol uptake under the high-rate denitrification process. In a sequencing batch reactor, the denitrifying activity was enriched up to 0.80 g-N/g-VSS-day for 72 days. Then, the enriched denitrifying sludge was transferred to a completely stirred tank reactor (CSTR). At the final phase on Day 46-50, the nitrogen removal efficiency was around 100% and the total nitrogen removal rate reached 0.097±0.003 kg-N/㎥-day. During the continuous process, the sludge settling index (SVI30) was stabilized as 118.3 mL/g with the biomass concentration of 1,607 mg/L. The continuous denitrifying process was accelerated by using a sequencing batch reactor (SBR) with a total nitrogen removal rate of 0.403±0.029 kg-N/㎥-day with a high biomass concentration of 8,433 mg-VSS/L. Because the reactor was open to ambient air with the dissolved oxygen range of 0.2-0.5 mg-O2/L, an increased organic carbon requirement of 5.58±0.70 COD/NO3--N was shown for the SBR in comparison to the value of 4.13±0.94 for the test of the same biomass in a completely anaerobic batch reactor. The molecular analysis based on the 16S rRNA gene showed that Methyloversatilis discipulorum and Hyphomicrobium zavarzinii were the responsible denitrifiers with the sole organic carbon source of methanol.

Bacterial Community of Free-living and Aggregated Bacteria at Thawing Period in Lake Baikal (해빙기 바이칼호에서 부유세균과 Aggregates에 부착한 세균의 군집구조)

  • 홍선희;김옥선;전선옥;유재준;안태석
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.192-197
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    • 2002
  • Fluorescent in situ hybridization (FISH) with rRNA-targeted oligonucleotide probes was used to compare the community structures of free-living and aggregated bacteria at thawing period in Lake Baikal. Targeted groups were Eubacteria, $\alpha$-, $\beta$-, $\gamma$- proteobacteria groups, Cytophaga-Flavobacterium group and Planctomycetales. Total bacterial numbers of free-living bacteria were ranged from $0.2{\times}10^6\cells{\cdot}ml^-1$ to $3.2{\times}10^6\cells{\cdot}ml^-1$, which were decreasing with depth, while the aggregated bacterial numbers were dramatically increasing from $0.4{\times}10^4 to 3.3{\times}10^4 \cells{\cdot}ml^-1$ with depth. The ratios of EUB probe binding cells to DAPI counts were ranged from 52.3 to 74.1% in free-living bacteria, and from 39.6 to 66.7% in the aggregated bacteria, respectively. Community structures of the aggregated bacteria were very different from each free-living bacteria at every depth. At 25 m depth, where the chlorophyll a concentration was highest, both structures were quite different from those of surface layers, rendering the fact that the community structures might be affected by phytoplankton. The vertical profile of community structure of aggregated bacteria is particular. The proportion of $\beta$-proteobacteria group was increasing with depth and it was 51.8% at 100 m, but the dominant group was $\gamma$-pro-teobacteria group at 250 m. Taken together, the biodiversity and succession of aggregated bacteria are quite different from free-living bacteria.

Analysis of Bacterial Community Structure of Biological Activated Carbon Process in Drinking Water Treatment Plant Using FISH (FISH법을 이용한 정수처리장 내 생물활성탄 공정의 세균군집 구조 분석)

  • Son, Hyeng-Sik;Kim, Mi-A;Jeong, Seong-Yun;Kim, Young-Hun;Son, Hee-Jong;Park, Geun-Tae;Kim, Min-Ju;Ryu, Eun-Yeon;Lee, Sang-Joon
    • Journal of Environmental Science International
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    • v.17 no.5
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    • pp.555-564
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    • 2008
  • The bacterial community structure in biological activated carbon (BAC) process in drinking water treatment plant was investigated by Fluorescent in situ Hybridization (FISH) with rRNA-targeted oligonucleotide probe. Samples were collected at different three points in BAC process every month for one year. They were hybridized with a probe specific for the alpha, beta, gamma subclass of the class Proteobacteria, Cytophaga-Flavobacteria group and Gram-positive high G+C content (HGC) group. Total numbers of bacteria in BAC process counted by 4',6-diamidino-2-phenylindole (DAPI) staining were $5.4{\times}10^{10}$ (top), $4.0{\times}10^{10}$ (middle) and $2.8{\times}10^{10}$ cells/ml (bottom). The number of the culturable bacteria was from $1.0{\times}10^7$ to $3.6{\times}10^7$ cells/ml and the culturability was about 0.05%. The faction of bacteria detectable by FISH with the probe EUB338 was about 83% of DAPI counts. Gamma and alpha subclass of the class Proteobacteria were predominant in BAC process and their ratios were over 20% respectively. In top and middle, alpha, beta and gamma subclass of the class Proteobacteria competed with each other and their percentages was changed according to the season. In bottom, gamma subclass of the class Proteobacteria was predominant all through the year. It could be successfully observed the seasonal distribution of bacterial community in biological activated carbon process using FISH.

The complete plastid genome and nuclear ribosomal transcription unit sequences of Spiraea prunifolia f. simpliciflora (Rosaceae)

  • Jeongjin CHOI;Wonhee KIM;Jee Young PARK;Jong-Soo KANG;Tae-Jin YANG
    • Korean Journal of Plant Taxonomy
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    • v.53 no.1
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    • pp.32-37
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    • 2023
  • Spiraea prunifolia f. simpliciflora Nakai is a perennial shrub widely used for horticultural and medicinal purposes. We simultaneously obtained the complete plastid genome (plastome) and nuclear ribosomal gene transcription units, 45S nuclear ribosomal DNA (nrDNA) and 5S nrDNA of S. prunifolia f. simpliciflora, using Illumina short-read data. The plastome is 155,984 bp in length with a canonical quadripartite structure consisting of 84,417 bp of a large single-copy region, 18,887 bp of a short single-copy region, and 26,340 bp of two inverted repeat regions. Overall, a total of 113 genes (79 protein-coding genes, 30 tRNAs, and four rRNAs) were annotated in the plastome. The 45S nrDNA transcription unit is 5,848 bp in length: 1,809 bp, 161 bp, and 3,397 bp for 18S, 5.8S, and 26S, respectively, and 261 bp and 220 bp for internal transcribed spacer (ITS) 1 and ITS 2 regions, respectively. The 5S nrDNA unit is 512 bp, including 121 bp of 5S rRNA and 391 bp of intergenic spacer regions. Phylogenetic analyses showed that the genus Spiraea was monophyletic and sister to the clade of Sibiraea angustata, Petrophytum caespitosum and Kelseya uniflora. Within the genus Spiraea, the sections Calospira and Spiraea were monophyletic, but the sect. Glomerati was nested within the sect. Chamaedryon. In the sect. Glomerati, S. prunifolia f. simpliciflora formed a subclade with S. media, and the subclade was sister to S. thunbergii and S. mongolica. The close relationship between S. prunifolia f. simpliciflora and S. media was also supported by the nrDNA phylogeny, indicating that the plastome and nrDNA sequences assembled in this study belong to the genus Spiraea. The newly reported complete plastome and nrDNA transcription unit sequences of S. prunifolia f. simpliciflora provide useful information for further phylogenetic and evolutionary studies of the genus Spiraea, as well as the family Rosaceae.

The study on the structure of the Lee Je-ma's viewpoint of Morpho-Image (이제마(李濟馬) 형상관(形象觀)의 구조적(構造的) 고찰(考察))

  • Lee, Jun-hee;Koh, Byung-hee;Song, Il-byung
    • Journal of Sasang Constitutional Medicine
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    • v.11 no.1
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    • pp.41-61
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    • 1999
  • The purpose of this article is to investigate Lee Je-rna'os viewpoint of Morpho-Image. In the first place, I will define the viewpoint of Morpho-Image as the the structure of recognition and study it through bibliographic study on Lee Je-ma's work. The conclusions summarized as follows; 1. The structure of recognition, represented in "Gyukchigo(格致藁)" "Dongyi Soose Bowon Sasang Chobonkyon(東醫壽世保元四象草本卷)", is based on Taigi(太極), Liangyi(兩儀), Sasang(四象). There are the mind of Taigi(太極), the mind of Liangyi(兩儀) separated from Taigi(太極), the mind of Sasang(四象) separated from Liangyi(兩儀). The mind of Taigi(太極) is the mind of center, the mind of Liangyi(兩儀) is the mind of the mind and body(or the wisdom and action). Two axis, the mind(or wisdom) and the body(or action), cross and make Sasang(四象), affair-mind-body-object which classifies all affairs and objects of humans, society, and the universe as four types. 2. Lee Je-ma summarized everything in view of Sasang(四象), Sasang(四象) in view of Liangyi(兩儀), Liangyi(兩儀) in view of Taigi(太極). 3. The procedure of recognition above is described in , , . Such is transformed in terms of human body and described in .

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EVALUATION OF OSTEOGENIC ACTIVITY AND MINERALIZATION OF CULTURED HUMAN DENTAL PAPILLA-DERIVED CELLS (배양된 치유두 유래세포의 조골활성 및 골기질 형성의 평가)

  • Park, Bong-Wook;Byun, June-Ho;Choi, Mun-Jeoung;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.4
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    • pp.279-288
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    • 2007
  • In the present study, we focused on stem cells in the dental papilla of the tooth germ. The tooth germ, sometimes called the tooth bud, is the primordial structure from which a tooth is formed. The tooth germ consists of the enamel organ, the dental papilla, and the dental follicle. The dental papilla lies below a cellular aggregation of the enamel organ. Mesenchymal cells within the dental papilla are responsible for formation of dentin and pulp of a tooth. Tooth germ disappears as a tooth is formed, but that of a third molar stays in the jawbone of a human until the age of 10 to 16, because third molars grow slowly. Impacted third molar tooth germs from young adults are sometimes extracted for orthodontic treatment. In the present study, we evaluated the osteogenic activity and mineralization of cultured human dental papilla-derived cells. Dental papillas were harvested from mandible during surgical extraction of lower impacted third molar from 3 patients aged 13-15 years. After passage 3, the dental papilla-derived cells were trypsinized and subsequently suspended in the osteogenic induction DMEM medium supplemented with 10% fetal bovine serum, 50 g/ml L-ascorbic acid 2-phosphate, 10 nM dexamethasone and 10 mM -glycerophosphate at a density of $1\;{\times}10^6\;cells/dish$ in a 100-mm culture dish. The dental papilla-derived cells were then cultured for 6 weeks and the medium was changes every 3 days during the incubation period. Dental papilla-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 7 of culture period, then decreased in intensity during the culture period. ALP mRNA level was largely elevated at 1 weeks and gradually decreased with culture time. Osteocalcin mRNA expression appeared at day 14 in culture, after that its expression continuously increased in a time-dependent manner up to day 28. The expression remained constant thereafter. Runx2 expression appeared at day 7 with no detection thereafter. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. Osteocalcin secretion was detectable in the culture medium from 1 week. The secretion of osteocalcin from dental papilla-derived cells into the medium greatly increased after 3 weeks although it showed a shallow increase by then. In conclusion, our study showed that cultured human dental papilla-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix.

Biological Functions of the COOH-Terminal Amino Acids of the $\alpha$-Subunit of Tethered Equine Chorionic Gonadotropin

  • Jeoung, Youn-Hee;Yoon, Jong-Taek;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.34 no.1
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    • pp.47-53
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    • 2010
  • Glycoprotein hormones have a common $\alpha$-subunit that is involved in the signaling pathway together with G protein, adenylcyclase and cAMP induction; however, it is an unclear how this common structure is related to hormonal action. To determine the biological functions of the COOH-terminal amino acids in the $\alpha$-subunit of these glycoprotein hormones, a tethered-molecule was constructed by fusing the $NH_2$-terminus of the $\alpha$-subunit to the COOH-terminus of the $\beta$-subunit of equine chorionic gonadotropin (eCG). The following deletion mutants were created by PCR; Ile was inserted at position 96 to form ${\Delta}96$, Lys was substituted at position 95 to form ${\Delta}95$, His was inserted at position 93 to form ${\Delta}93$ and Tyr was substituted at position 87 to form ${\Delta}87$. Each mutant was transfected into CHO-K1 cells. Tethered-wt eCG, and ${\Delta}96$, ${\Delta}95$, and ${\Delta}93$ mutants were efficiently secreted into the medium but the ${\Delta}87$ mutant was not secreted. Interestingly, the RT-PCR, real-time PCR, and northern blot analyses confirmed that the RNA was transcribed in the ${\Delta}87$ mutant. However, the ${\Delta}87$ mutant protein was not detected in the medium or the intracellular fraction of the cell lysates. The LH- and FSH-like activities of the recombinant proteins were assayed in terms of cAMP production using rat LH/CG and rat FSH receptors. The metabolic clearance rate (MCR) was determined by injecting rec-eCG (2 IU) into the tail vein. The ${\Delta}95$ and ${\Delta}93$ mutants were completely inactive in both the LH- and FSH-like activity assays. The ${\Delta}96$ mutant showed slight activity in the LH-like activity assay. In comparison to the wild type, the activity of the ${\Delta}96$ mutant in the FSH-like activity assay was the highest among all the mutants. The MCR assay in which rec-eCG was injected showed a peak at 10 min in all the treatment groups, which disappeared 4 h after injection. These results imply a direct interaction between the receptor and the COOH-terminal region of the a-subunit. The data also reveal a significant difference in the mechanism by which the eCG hormone interacts with the rLH and rFSH receptors. The COOH-terminal region of the $\alpha$-subunit is very important for the secretion and functioning of this hormone.

Bacterial core community in soybean rhizosphere (콩 근권의 핵심 세균 군집)

  • Lee, Youngmi;Ahn, Jae-Hyung;Choi, Yu-Mi;Weon, Hang-Yeon;Yoon, Jung-Hoon;Song, Jaekyeong
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.347-354
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    • 2015
  • Soybean is well known to be originated from Korea and far-east Asian countries, and studies of many root nodule bacteria associated with soybean have mainly-focused on nitrogen fixation, but much less study was carried out on bacterial community in the rhizosphere of soybean. In this study, we analyzed the bacterial community in rhizosphere of Korean soybean, Daepungkong using the pyrosequencing method based on the 16S rRNA gene to characterize the change of the rhizosphere community structure according to the growth stages of soybeans and to elucidate bacterial core community in rhizosphere of soybean. Our results revealed that bacterial community of rhizosphere soil differed from that of bulk soil and was composed of a total of 21 bacterial phyla. The predominant phylum in the rhizosphere of soybean was Proteobacteria (36.6-42.5%) and followed by Acidobacteria (8.6-9.4%), Bacteroidetes (6.1-10.9%), Actinobacteria (6.4-9.8%), and Firmicutes (5.7-6.3%). The bacterial core community in soybean rhizosphere was mainly composed of the operational taxonomic units (OTUs) belonging to the phylum Proteobacteria throughout all growth stages. The OTU00006 belonged to the genus Bradyrhizobium had the highest abundance and Steroidobacter, Streptomyces, Devosia were followed. These results show that bacterial core community in soybean rhizosphere was mainly composed of OTUs associated with plant growth promotion and nutrient cycles.

HeLa Cell Culture on Nanoimprinted Patterns Using Conducting Polymer (전도성 고분자 나노임프린트 패턴 상의 HeLa 세포 배양)

  • Ahn, Junhyoung;Park, Kyungsook;Lee, Suok;Jung, Sanghee;Lim, Hyungjun;Shin, Yong-Beom;Lee, JaeJong
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.41 no.1
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    • pp.63-67
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    • 2017
  • In bioscience and biotechnology, the research of fundamental life mechanisms and their diseases caused by insufficiency is important. The study of a whole organism is difficult and sometimes impossible because of DNA, RNA, proteins, cellular organelles, various cells, and organs. Cell cultures can provide a simple method for researching cellular mechanisms and conditions, both in terms of physiological performance, and in response to chemical stimulation. According to conventional cell culture methodology, the flat surface is used with surface treatments for cell adhesion on the surface. Micro- and nanoscale patterns have been developed with chemical and biochemical modifications for cell immobilization. In this study, HeLa cell culture on nanostructures patterns was studied, including the 300 nm line and 150 nm pillar structures, using nanoimprint lithography and pyrrole as a biocompatible conducting polymer.