• Title/Summary/Keyword: molecular analyses

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Molecular Genetic Classification of Hypsizigus marmoreus and Development of Strain-specific DNA Markers (느티만가닥버섯의 분자유전학적 분류 및 품종특이적 DNA 마커 탐색)

  • Lim, Yun-Jeong;Lee, Chang-Yun;Park, Jeong-Eun;Kim, Sang-Woo;Lee, Hyun-Sook;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.34-39
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    • 2010
  • We have attempted to verify 30 strains of Hypsizigus marmoreus from various mushroom stocks in Korea using random amplified polymorphic DNA (RAPD) methodology. Chromosomal DNAs of them were extracted and subjected to PCR analyses with 3 random primers. Each PCR produced approximately 30 distinct PCR bands with the size from 200 bp to 3000 bp. A dendrogram was acquired using the unweighted pair-group method with arithmetic average (UPGMA) clustering methodology on the basis of the DNA band pattern. The analysis revealed that 30 strains of H. marmoreus were clustered into two distinct clusters. Cluster 1 contained 3 subgroups while the cluster 2 consisted of rather diverse strains. Interestingly, Hm3-10, a wild strain collected from Deog-Yu mountain, was not included in either clusters, indicative of uniqueness of this strain. We nextly attempted to develop strain-specific DNA markers to verify a specific strain. A unique band in the RAPD gel lane of Hm0-4 was extracted and its sequence was determined. PCR with a primer set from the determined sequence revealed that the primer set gave a 250 bp DNA band only for Hm0-4, indicating that this approach works well for the strain-specific identification of H. marmoreus.

Multi-locus Phylogeny Analysis of Korean Isolates of Phytophthora Species Based on Sequence of Ribosomal and Mitochondrial DNA (핵 및 미토콘드리아 DNA 염기서열을 이용한 국내 Phytophthora 속의 Multi-locus phylogeny 분석)

  • Seo, Mun-Won;Song, Jeong-Young;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.40-47
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    • 2010
  • To investigate genetic relationships either interspecies or intraspecies of 14 Korean Phytophthora species, sequence analyses of nuclear DNA (ypt gene and rDNA-IGS region) and mitochondrial DNA (Cox gene, $\beta$-tubuline gene, and EF1A gene) were performed. All of 14 Korean Phytophthora species clearly clustered into foreign isolates of each species. These Korean isolates in Phytophthora species also showed no correlation between molecular classification and morphological classification like as in case of foreigners. P. palmivora KACC 40167 reported previously from genetic groups of Phytophthora species in Korea was not consistent with the classification system, and therefore was required re-examination for the genetic group analysis. Korean isolates of P. drechsleri KACC 40195 showed very close relationship with P. cryptogea KACC 40161 above 94% bootstrap value in P. cryptogea-P. drechsleri complex group. Identification of these isolates is still unclear, because P. cryptogea and P. drechsleri were not differentiated in this study. On the other hand, it was required to unify species for these two species, since P. parasitica and P. nicotianae were clustered into a group on the level of 99 to 100% sequence homology. Comparing to the sequences of foreigners, Korean isolates were newly divided to ten groups in the phylogenic system. These results could be prepared useful informations to understand genetic diversity of Phytophthora species in Korea.

Miscibility of Melt-mixed PLLA/PMMA Blends for Optical Film Application (광학 필름 적용을 위한 용융혼합된 PLLA/PMMA 블렌드의 상용성 연구)

  • Park, Eun Ju;Kim, In Seok;Park, Sang Seok;Lee, Ho Sang;Lee, Moo Sung
    • Polymer(Korea)
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    • v.37 no.6
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    • pp.744-752
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    • 2013
  • The miscibility between poly(L-lactide) (PLLA) and poly(methyl methacrylate) (PMMA) was investigated using thermal analyses for the purpose of developing birefringence-free material at oriented state. The effect of methyl acrylate (MA) units as comonomer of PMMA on the miscibility was also studied. All the blends prepared in this study show composition-dependent single $T_g$'s between those of blend components and high transparency over the visible region, indicating the miscibility at molecular level and no discernible effect of MA units on it. No phase separation was observed at elevated temperature of $280^{\circ}C$, higher than the degradation of PLLA and PMMA. The interaction energy density in PLLA/PMMA blends with 17 mol% of MA was measured to be $-0.74J/cm^3$ from the equilibrium melting temperature depression based on the Hoffman-Weeks method. The blends show zero-${\Delta}$n behavior at a specific mixing ratio and the drawing ratio of 3 due to compensation of intrinsic orientation birefringence. Birefringence dispersion of PLLA/PMMA5 blends was also measured to examine the possibility for quarter-wave plates or polarizer protective films.

Germ Line Transformation of the Silkworm, Bombyx mori L. with a piggyBac Vector Harboring the Human Lactoferrin Gene (락토페린 유전자도입 piggyBac 벡터에 의한 누에 형질전환)

  • Kim, Yong-Soon;Sohn, Bong-Hee;Kim, Kee-Young;Jung, I-Yeon;Kim, Mi-Ja;Kang, Pil-Don
    • Journal of Sericultural and Entomological Science
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    • v.49 no.2
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    • pp.37-42
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    • 2007
  • Lactoferrin, an ion-binding 80-kDa glycoprotein, has been suggested to have many biologic activities, such as facilitating ion absorption and having antimicrobial and anti-inflammatory effects. Several of these activities are likely to only be facilitated by human lactoferrin because they depend on the binding of human lactoferrin to specific receptor. To produce recombinant human lactoferrin to animal foods using transgenic silkworm, Bombyx mori L, we have cloned and sequenced the cDNA encoding for a human lactoferrin (HLf) from the mRNA in mammary tumor line (GI-101). As a result, the 2.5-kb fragment of HLf gene was cloned with pGEM-T vector and then this fragment was sequenced. In the nucleotide sequence analysis, single open reading frame of the 2,136-bp encoding for a polypeptide of 712 amino acid residues was detected. On the other hand, we constructed a recombinant plasmid(pPT-HLf), containing human lactoferrin gene for germ line transformation of the silkworm using a piggyBac transposon-derived vector. A nonautonomous helper plasmid encodes the piggyBac transposase. Approximately 6.7% of individuals in the G0 silkworms expressed green fluorescent protein (GFP). PCR analyses of GFP-positive silkworms (G0 and G1) revealed that independent insertions occurred frequently. Furthermore, Western blot analysis showed that the recombinant HLf expressed in hemolymph has the same molecular weight (80 kDa) as a native protein. On the basis of these experiments, expression of HLf in next generation of transgenic silkworm is now in process.

Vegetation Structure and Management Strategies of Glaux maritima var. obtusifolia Community on the Southernmost Distribution Area in Korea (멸종위기 식물인 갯봄맞이 최남단 군락의 식생구조)

  • Lim, Jeong Cheol;Lee, Cheol Ho;Kim, Eui Ju;Choi, Byoung Ki
    • Journal of Wetlands Research
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    • v.20 no.1
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    • pp.1-13
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    • 2018
  • Glaux maritima var. obtusifolia is distributed in very limited areas in South Korea and thus designated and protected as an endangered species. This study aimed to identify the diversity of vegetation in the Dangsa coast wetlands, the southern limit of G. maritima var. obtusifolia and to assess determinants of the vegetation and the importance of habitats. The phytosociological method of the $Z{\ddot{u}}rich-Montpellier$ School was used for vegetation classification and mathematical and statistical analyses were also conducted to analyze environmental factors and their relationship with the vegetation. The results of this study showed that there were 4 vegetation units in the Dangsa coast wetlands: Glaux maritima var. obtusifolia-Triglochin maritimum community (included three subcommunities), Puccinellia nipponica s.l. community, Beckmannia syzigachne-Isachne globosa community and Typha laxmannii-Phragmites communis community. It was also found that major determinants of the vegetation include moisture environment, soil depth, water level disturbance, vegetation height, community structure, etc. Glaux maritima was identified to grow most dominantly in the typicum subassociation of Glaux. maritima var. obtusifolia-Triglochin maritimum community, and the species compositions and dominant situations were observed to be similar to those in the southern limit in Japan, adjacent to South Korea. The assessment results indicated that the Dangsa coastal wetlands have a significant meaning from phytogeographical and syngeographical aspects, and contribute as a shelter for diverse species. It is required to establish conservation strategies to accurately determine the value of the wetlands of the Dangsa coast from various perspectives, and to protect and manage them.

Immuno-stimulating Polysaccharides from the Fruiting Bodies of Fomitella fraxinea (I) - Characterization of polysaccharides extracted with neutral sodium chloride solution - (Fomitella fraxinea로부터 분리한 면역활성 다당류 (I) -중성염 용액 추출 다당류의 특성-)

  • Cho, Soo-Muk;Lee, Jae-Hoon;Han, Sang-Bae;Kim, Hwan-Mook;Yu, Seung-Hun;Yoo, Ick-Dong
    • The Korean Journal of Mycology
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    • v.23 no.4 s.75
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    • pp.332-339
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    • 1995
  • Water-soluble polysaccharide (FCW) was extracted from the fruiting body of Fomitella fraxinea with neutral sodium chloride solution. The polysaccharide was further fractionated into FCW-I and FCW-II by ion exchange chromatography. The FCW-I and FCW-II were then purified by gel permeation chromatography and named as FCW-Ia and FCW-IIa, respectively. FCW-IIa showed relatively strong immuno-stimulating activity but FCW-Ia did not. By analyses of HPLC and GPC, FCW-Ia and FCW-IIa were identified to be homogeneous and their molecular weights were estimated to be about 15,000 and 8,700, respectively. FCW-Ia consisted of fucose, galactose, and mannose as main sugars and their molar ratio was 19.5 : 63.2 : 25.0. Protein was not detected in FCW-Ia. However, FCW-IIa was composed of glucose, galactose, and mannose at a molar ratio of 1.0 : 0.3 : 0.4 and contained 0.4% protein with a higher amount of glutamic acid. A small amount of uronic acid was detected in both FCW-Ia and FCW-IIa.

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Prevalence, Associated Risk Factors, and Phylogenetic Analysis of Toxocara vitulorum Infection in Yaks on the Qinghai Tibetan Plateau, China

  • Li, Kun;Lan, Yanfang;Luo, Houqiang;Zhang, Hui;Liu, Dongyu;Zhang, Lihong;Gui, Rui;Wang, Lei;Shahzad, Muhammad;Sizhu, Suolang;Li, Jiakui;Chamba, Yangzom
    • Parasites, Hosts and Diseases
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    • v.54 no.5
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    • pp.645-652
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    • 2016
  • Toxocara vitulorum has been rarely reported in yaks at high altitudes and remote areas of Sichuan Province of Tibetan Plateau of China. The current study was designed to investigate the prevalence, associated risk factors, and phylogenetic characteristics of T. vitulorum in yak calves on the Qinghai Tibetan plateau. Fecal samples were collected from 891 yak calves and were examined for the presence of T. vitulorum eggs by the McMaster technique. A multivariable logistic regression model was employed to explore variables potentially associated with exposure to T. vitulorum infection. T. vitulorum specimens were collected from the feces of yaks in Hongyuan of Sichuan Province, China. DNA was extracted from ascaris. After PCR amplification, the sequencing of ND1 gene was carried out and phylogenetic analyses was performed by MEGA 6.0 software. The results showed that 64 (20.1%; 95% CI 15.8-24.9%), 75 (17.2; 13.8-21.1), 29 (40.9; 29.3-53.2), and 5 (7.6; 2.5-16.8) yak calves were detected out to excrete T. vitulorum eggs in yak calve feces in Qinghai, Tibet, Sichuan, and Gansu, respectively. The present study revealed that high infection and mortality by T. vitulorum is wildly spread on the Qinghai Tibetan plateau, China by fecal examination. Geographical origin, ages, and fecal consistencies are the risk factors associated with T. vitulorum prevalence by logistic regression analysis. Molecular detection and phylogenetic analysis of ND1 gene of T. vitulorum indicated that T. vitulorum in the yak calves on the Qinghai Tibetan plateau are homologous to preveiously studies reported.

Screening and Identification of Soy Curd-Producing Lactic Acid Bacteria (두유 커드를 생산하는 김치 유래 젖산균의 동정)

  • Kim, Ro-Ui;Ahn, Soon-Cheol;Yu, Sun-Nyoung;Kim, Kwang-Youn;Seong, Jong-Hwan;Lee, Young-Guen;Kim, Han-Soo;Kim, Dong-Seob
    • Journal of Life Science
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    • v.21 no.2
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    • pp.235-241
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    • 2011
  • The purpose of this study was to isolate soy curd forming bacterial strains. Soy curd forming bacteria were isolated from Kimchi, a traditional Korean vegetable food that is fermented using lactic acid bacteria. Among 196 bacterial strains, ten isolates (strain No. 2-2-2, 2-15-2, 2-18-1, 2-19-2, 3-4-1, 3-4-2, 3-8-1, 3-8-3, 3-17-1, 4-39-5) formed firm soy curd. The isolated bacterial strains were identified by molecular biological and biochemical analyses. The genomic DNAs extracted from the isolated bacterial strains were used as a template for PCR amplification of 16S rDNA region. By comparing the results of the 16s rDNA sequences with GenBank data, the isolated strains were identified as Leuconostoc mesenteroides group and Lactobacillus sakei group. The phylogenetic position of soy curd forming strains and their related taxa were investigated using neighbor-joining method. L. mesenteroides group was further identified as L. mesenteroides subsp. dextranicum based on biochemical properties. L. sakei group was named Lactobacillus sp., because it showed a variety of biochemical properties.

Smad-dependent Expression of Gadd45b Gene during TGF-β-induced Apoptosis in EpH4 Cells. (EpH4 세포에서 TGF-β에 의한 세포사멸시 Smad 단백질에 의존한 Gadd45b 유전자의 발현 변화)

  • Cho, Hee-Jun;Yoo, Ji-Yun
    • Journal of Life Science
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    • v.18 no.4
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    • pp.461-466
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    • 2008
  • Transforming growth $factor-{\beta}$ ($TGF-{\beta}$)-dependent apoptosis is important in the elimination of damaged or abnormal cells from normal tissues in vivo. Gadd45b has been known to participate in $TGF-{\beta}-induced$ apoptosis by the activation of p38 kinase. In this report, we show that Gadd45b is an immediate-early response gene for $TGF-{\beta}$ during apoptosis in EpH4 cells. To elucidate the molecular mechanism of $TGF-{\beta}-induced$ Gadd45b gene expression, we cloned the 5'-flanking region of the mouse Gadd45b gene. When transfected into EpH4 cells, this 5'-flanking region conferred promoter activity and inducibility by $TGF-{\beta}$. Deletion analyses demonstrated that the minimal promoter activity was detected in the proximal region 220 bp upstream of the transcription initiation site. We also found that the proximal Gadd45b promoter is activated by $TGF-{\beta}$ through the action of Smad2, Smad3, and Smad4. Finally, we show that the expression of Gadd45b gene by $TGF-{\beta}$ is suppressed in EpRas cells in which $TGF-{\beta}$ could not induce apoptosis, suggesting that Gadd45b may be a crucial target for $TGF-{\beta}-induced$ apoptosis in EpH4 cells.

Synergistic Effects and Mechanism of Coal/Polypropylene Coliquefaction (석탄과 폴리프로필렌의 공동액화시 상승효과 및 반응기구)

  • Jeong, Daeheui;Kim, Seungrok;Lee, Jinyoung;Jeong, Taejin;Han, Choon
    • Applied Chemistry for Engineering
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    • v.9 no.5
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    • pp.742-748
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    • 1998
  • Experiments have been conducted to investigate synergistic effects and mechanisms of the Alaskan subbituminous coal/polypropylene (PP). Coliquefaction of coal/PP gave the synergistic effect in yields by 17% compared to independent liquefactions of coal or PP at $430^{\circ}C$. To analyse coliquefaction mechanisms, the second and zeroth order kinetic models were developed for coal and PP conversions respectively. When the models were simulated to fit coliquefaction results, those represented results successfully with the correlation coefficient of 0.99. The amount of tetralin needed to liquefy unit mass of coa 1(${\beta}$) was also calculated using the developed model. According to the calculated results, $\beta$ increased from 0.4 to 1.0 as the coliquefaction temperature increased from $410^{\circ}C$ to $470^{\circ}C$. This indicated that tetralin lowered the molecular weight of oil produced, and this phenomenon was recognized by the GPC analyses. Furthermore, it was found that tetralin prohibited the liquefaction of PP when coal was not added. On the other hand, tetralin was believed to act as a hydrogen-donor solvent to coal rather than prohibit PP liquefaction during coliquefaction. Therefore, the liquefaction rate of PP increases and synergistic effects in oil yields are mainly due to the increase in PP liquefaction during coal/PP coliquefaction.

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