• Title/Summary/Keyword: molecular species

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Analysis of Potential Toxigenicity and Phylogeny using Target Genes in Aphanizomenon flos-aquae (Cyanophyceae) strains isolated from the Nakdong River (낙동강에서 분리된 Aphanizomenon flos-aquae (Cyanophyceae) 균주의 목표 유전자를 이용한 잠재적 독소 생성능 및 계통학적 분석)

  • Ryu, Hui-Seong;An, Sung-Min;Lim, Chang-Kun;Shin, Ra-Young;Park, Jong-Guen;Lee, Jung-Ho
    • Korean Journal of Ecology and Environment
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    • v.50 no.1
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    • pp.137-147
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    • 2017
  • The identity of toxin producers remains only hypothesis unless there were identified by strain isolation and analytical confirmation of both the cyanotoxin production and the genetic identity of the monoculture. The purposes of this study were to identify a morphologic and phylogenetic classification in Aphanizomenon flos-aquae strains isolated from the Nakdong River and to investigate the potential ability of the strains to produce toxins such as saxitoxin and cylindrospermopsin using target genes. The 16S rRNA and sxtA, sxtI, cyrA, cyrJ genes were analyzed on two strains (DGUC001, DGUC003) isolated from the Nakdong River. Morphological features of the strains were observed a shape of aggregated trichomes in parallel fascicles which can reach up to macroscopic size and a hyaline terminal cell without aerotope. In addition, the 16S rRNA phylogenetic analyses showed that the strains were identified as the same species with high genetic similarity of 98.4% and grouped within a monospecific andsupported cluster I of Aphanizomenon flos-aquae selected from GenBank of the NCBI. The cyrA and cyrJ genes encoding for the cylindrospermopsin-biosynthesis were not detected in the present study. The sxtA gene was in detected both the two strains, whereas the sxtI gene which had been suggested as a suitable molecular marker to detect saxitoxin-producing cyanobacteria was not found both the strains. Thus, the two strains isolated from Nakdong River were identified as the same species of Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault 1888, the two strains were confirmed as potential non-producing strains of the saxitoxin and cylindrospermopsin.

Mechanisms for Anti-wrinkle Activities from Fractions of Black Chokeberries (블랙초크베리 분획물로부터의 주름억제 효과에 대한 작용기전)

  • Choi, Eun-Young;Kim, Eun-Hee;Lee, Jae-Bong;Do, Eun-Ju;Kim, Sang-Jin;Kim, Se-Hyeon;Park, Jeong-Yeol;Lee, Jin-Tae
    • Journal of Life Science
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    • v.26 no.1
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    • pp.34-41
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    • 2016
  • Black chokeberries (scientific name Aronia melanocarpa) have been reported to have major effects due to anti-oxidant, anti-inflammatory, and anti-cancer capabilities. In this study, we investigated the anti- wrinkle effects of A. melanocarpa, including collagenase inhibition effects and their molecular biological mechanisms, such as oxidative stress-induced matrix metalloproteinase (MMP), mitogen-activated protein (MAP) kinase, and activator protein (AP)-1 expression and/or phosphorylation. In collagenase inhibition activity, the ethyl acetate fraction of black chokeberry (AE) was 77.2% at a concentration of 500 μg/ml, which was a significant result compared to that of Epigallocatechin gallate (positive control, 83.9% in 500 μg/ml). In the reactive oxygen species (ROS) assay, the AE produced 78% of ROS in 10 μg/ml and 70% of ROS in 75 μg/ml, which was a much lower percentage than the ROS production of H2O2-induced CCRF S-180II cells. In the MTT assay, cell viability was increased dose-dependently with AE in H2O2-induced cells. In protein expression by western blot assay, the AE suppressed the expression and phosphorylation of MMPs (MMP-1, -3, -9), MAPK (ERK, JNK, and p38), and AP-1 (c-Fos and c-Jun), and expressed the pro-collagen type I in H2O2-induced cells. These results suggest that black chokeberries have anti-wrinkle and collagen-production effects, and they may be used in applications for material development in the functional food and cosmetic industries.

Production of Violacein by a Novel Bacterium, Massilia sp. EP15224 Strain (Violacein을 생산하는 Massilia sp. EP15224 균주)

  • Yoon, Sang-Hong;Baek, Hee-Jin;Kwon, Soon-Wu;Lee, Chang-Muk;Sim, Joon-Soo;Hahn, Bum-Soo;Koo, Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.317-323
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    • 2014
  • Violacein has received much attention due to its various important biological activities, including broad-spectrum antibacterial and antifungal activity, anti-malarial, anti-tumoral, anti-oxidant, and anti-diarrheal activities. EP15224 strain isolated from forest soils in Korea was found to be a new species belonged to the genus Massilia based on its 16S ribosomal DNA sequences. The 16S ribosomal DNA of strain EP15224 displayed 97% homology with Massilia sp. BS-1, the nearest violacein-producing bacterium. Strain EP15224 produced bluish-purple pigment well in a synthetic MM2 medium containing glucose, $(NH_4)_2SO_4$, $Na_2HPO_4{\cdot}7H_2O$, $KH_2PO_4$, $MgSO_4{\cdot}7H_2O$, and 1 mM $\small{L}$-tryptophan. The chemical analysis of the pigment by LC/MS/MS showed that it is violacein with molecular weight of 343.34. This is the second report on the production of violacein by a Massilia species. In this study, the optimal culture conditions for violacein production were established under which 280 mg/l crude violacein was produced : glucose 2 g/l, $(NH_4)_2SO_4$ 1 g/l, $Na_2HPO_4{\cdot}7H_2O$ 2 g/l, $KH_2PO_4$ 1 g/l, $MgSO_4{\cdot}7H_2O$ 0.1 g/l, L-tryptophan 0.24 g/l, 25 ml medium in a 250 ml flask, with an inoculumn size of 10% (v/v), 72 h of cultivation with 250 rpm at $25^{\circ}C$.

Analyses of Genetic Relationships of Rhizoctonia solani Isolates from Various Crop Species and Rapid Identification of Anastomosis Groups with RAPD Method (각종 작물에서 분리한 R. solani 균주들의 RAPD를 이용한 종내 그룹의 유전적 유연관계 분석 및 AGs 신속 간이동정)

  • Lee, Youn-Su;Choi, Hei-Sun;Kim, Kyoung-Su;Woo, Su-Jin;Kang, Won-Hee;Kim, Myoung-Jo;Shim, Jae-Ouk;Lee, Min-Woong
    • The Korean Journal of Mycology
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    • v.26 no.3 s.86
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    • pp.373-379
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    • 1998
  • Rhizoctonia solani [Thanatephorus cucumeris (frank) Donk], one of the major soil-borne plant pathogens with world-wide distribution, can cause great damages on various crops. In Korea, sheath blight on rice caused by this pathogen is the major concern, and active studies on this pathogen have been performed. However, most of these studies were concerned with pathogenicity of the isolates instead of molecular analyses of different AGs of R. solani. Therefore, in this study, thirty isolates of Rhizoctonia solani collected from various sources were used for the analyses of genetic relationships among themselves and for the rapid anastomosis grouping with RAPD method. As a result, thirty isolates of known and unknown AGs were grouped into five subgroups and each group included AG-1, AG-2, AG-3, AG-4, and AG-5. RS-1 isolate was found to be closely related to AG-5. Isolates RS-4, RS-14, RS-17, and RS-16 were found to be closely related to AG-2-2(III B). Isolate RS-13 was closely related to AG-4, isolates RS-8 and RS-10 were closely related to AG-1(I B), and isolates RS-7 and RS-21 were closely related to AG-2-2(IV). Isolate RS-19 was closely related to AG-1(I C), and isolates RS-3, RS-5, RS-18, RS-6, and RS-15 were found to be closely related to AG-1.

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Relationship between Reactive Oxygen Species and Adenosine Monophosphate-activated Protein Kinase Signaling in Apoptosis Induction of Human Breast Adenocarcinoma MDA-MB-231 Cells by Ethanol Extract of Citrus unshiu Peel (진피 추출물에 의한 인간유방암 MDA-MB-231 세포의 apoptosis 유도에서 ROS 및 AMPK의 역할)

  • Kim, Min Yeong;HwangBo, Hyun;Ji, Seon Yeong;Hong, Su-Hyun;Choi, Sung Hyun;Kim, Sung Ok;Park, Cheol;Choi, Yung Hyun
    • Journal of Life Science
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    • v.29 no.4
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    • pp.410-420
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    • 2019
  • Citrus unshiu peel extracts possess a variety of beneficial effects, and studies on their anticancer activity have been reported. However, the exact mechanisms underlying this activity remain unclear. In the current study, the apoptotic effect of ethanol extract of C. unshiu peel (EECU) on human breast adenocarcinoma MDA-MB-231 cells and related mechanisms were investigated. The results showed that the survival rate of MDA-MB-231 cells treated with EECU was significantly inhibited in a concentration-dependent manner, which was associated with the induction of apoptosis. EECU-induced apoptosis was associated with the activation of caspase-8 and caspase-9, which initiate extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3, a representative effect caspase. EECU suppressed the expression of the inhibitor of apoptosis family of proteins, leading to an increased Bax/Bcl-2 ratio and proteolytic degradation of poly (ADP-ribose) polymerase. EECU also enhanced the loss of the mitochondrial membrane potential and cytochrome c release from the mitochondria to the cytosol, along with truncation of Bid. In addition, EECU activated AMP-activated protein kinase (AMPK), and compound C, an AMPK inhibitor, significantly weakened EECU-induced apoptosis and cell viability reduction. Furthermore, EECU promoted the generation of reactive oxygen species (ROS), which acted as upstream signals for AMPK activation as pretreatment of cells, with the antioxidant N-acetyl cysteine reversing both EECU-induced AMPK activation and apoptosis. Collectively, these findings suggest that EECU inhibits MDA-MB-231 adenocarcinoma cell proliferation by activating intrinsic and extrinsic apoptotic pathways, which was mediated through ROS/AMPK-dependent pathways.

Distribution and Frequency of SSR Motifs in the Chrysanthemum SSR-enriched Library through 454 Pyrosequencing Technology (국화 SSR-enriched library에서 SSR 반복염기의 분포 및 빈도)

  • Moe, Kyaw Thu;Ra, Sang-Bog;Lee, Gi-An;Lee, Myung-Chul;Park, Ha-Seung;Kim, Dong-Chan;Lee, Cheol-Hwi;Choi, Hyun-Gu;Jeon, Nak-Beom;Choi, Byung-Jun;Jung, Ji-Youn;Lee, Kyu-Min;Park, Yong-Jin
    • Journal of the Korean Society of International Agriculture
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    • v.23 no.5
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    • pp.546-551
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    • 2011
  • Chrysanthemums, often called mums or chrysanths, belong to the genus Chrysanthemum, which includes about 30 species of perennial flowering plants in the family Asteraceae. We extracted DNA from Dendranthema grandiflorum ('Smileball') to construct a simple sequence repeat (SSR)-enriched library, using a modified biotin-streptavidin capture method. GS FLX (Genome Sequencer FLX System which provides the flexibility to perform the broad range of applications) sequencing (at the 1/8 run specification) resulted in 18.83 mega base pairs (Mbp) with an average read length of 280.06 bp. Sequence analyses of all SSR-containing clones revealed a predominance of di-nucleotide motifs (16,375, 61.5%) followed by tri-nucleotide motifs (6,616, 24.8%), tetra-nucleotide motifs (1,674, 6.3%), penta-nucleotide motifs (1,283, 4.8%), and hexa-nucleotide motifs (693, 2.6%). Among the di-nucleotide motifs, the AC/CA class was the most frequently identified (93.5% of all di-nucleotide types), followed by the GA/AG class (6.1%), the AT/TA class (0.4%), and the CG/GC class (0.03%). When we analyzed the distribution of different repeat motifs and their respective numbers of repeats, regardless of the motif class, of 100 SSR markers, we found a higher number of di-nucleotide motifs with 70 to 80 repeats; we also found two di-nucleotide motifs with 83 and 89 repeats, respectively, but their product lengths were within optimum size (297 and 300 bp). In future work, we will screen for polymorphisms of possible primer pairs. The results will provide a useful tool for assessing molecular diversity and investigating the population structure among and within Chrysanthemum species.

A Basic Study on the Establishment of Preservation and Management for Natural Monument(No.374) Pyeongdae-ri Torreya nucifera forest of Jeju (천연기념물 제374호 제주 평대리 비자나무 숲의 보존·관리방향 설정을 위한 기초연구)

  • Lee, Won-Ho;Kim, Dong-Hyun;Kim, Jae-Ung;Oh, Hae-Sung;Choi, Byung-Ki;Lee, Jong-Sung
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.32 no.1
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    • pp.93-106
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    • 2014
  • In this study, Analyze environment of location, investigation into vegetation resources, survey management status and establish to classify the management area for Natural monument No.374 Pyengdae-ri Torreya nucifera forest. The results were as follows: First, Torreya nucifera forest is concerned about influence of development caused by utilization of land changes to agricultural region. Thus, establish to preservation management plan for preservation of prototypical and should be excluded development activity to cause the change of terrain that Gotjawal in the Torreya nucifera forest is factor of base for generating species diversity. Secondly, Torreya nucifera forest summarized as 402 taxa composed 91 familly 263 genus, 353 species, 41 varieties and 8 forms. The distribution of plants for the first grade & second grade appear of endangered plant to Ministry of Environment specify. But, critically endangered in forest by changes in habitat, diseases and illegal overcatching. Therefore, when establishing forest management plan should be considered for put priority on protection. Thirdly, Torreya nucifera representing the upper layer of the vegetation structure. But, old tree oriented management and conservation strategy result in poor age structure. Furthermore, desiccation of forest on artificial management and decline in Torreya nucifera habitat on ecological succession can indicate a problem in forest. Therefore, establish plan such as regulation of population density and sapling tree proliferation for sustainable characteristics of the Torreya nucifera forest. Fourth, Appear to damaged of trails caused by use. Especially, Scoria way occurs a lot of damaged and higher than the share ratio of each section. Therefore, share ratio reduction Plan should be considered through the additional development of tourism routes rather than the replacement of Scoria. Fifth, Representing high preference of the Torreya nucifera forest tourist factor confirmed the plant elements. It is sensitive to usage pressure. And requires continuous monitoring by characteristic of Non-permanent. In addition, need an additional plan such as additional development of tourism elements and active utilizing an element of high preference. Sixth, Strength of protected should be differently accordance with importance. First grade area have to maintenance of plant population and natural habitats. Set the direction of the management. Second grade areas focus on annual regeneration of the forest. Third grade area should be utilized demonstration forest or set to the area for proliferate sapling. Fourth grade areas require the introduced of partial rest system that disturbance are often found in proper vegetation. Fifth grade area appropriate to the service area for promoting tourism by utilizing natural resources in Torreya nucifera forest. Furthermore, installation of a buffer zone in relatively low ratings area and periodic monitoring to the improvement of edge effect that adjacent areas of different class.

A Novel Human BTB-kelch Protein KLHL31, Strongly Expressed in Muscle and Heart, Inhibits Transcriptional Activities of TRE and SRE

  • Yu, Weishi;Li, Yongqing;Zhou, Xijin;Deng, Yun;Wang, Zequn;Yuan, Wuzhou;Li, Dali;Zhu, Chuanbing;Zhao, Xueying;Mo, Xiaoyang;Huang, Wen;Luo, Na;Yan, Yan;Ocorr, Karen;Bodmer, Rolf;Wang, Yuequn;Wu, Xiushan
    • Molecules and Cells
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    • v.26 no.5
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    • pp.443-453
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    • 2008
  • The Bric-a-brac, Tramtrack, Broad-complex (BTB) domain is a protein-protein interaction domain that is found in many zinc finger transcription factors. BTB containing proteins play important roles in a variety of cellular functions including regulation of transcription, regulation of the cytoskeleton, protein ubiquitination, angiogenesis, and apoptosis. Here, we report the cloning and characterization of a novel human gene, KLHL31, from a human embryonic heart cDNA library. The cDNA of KLHL31 is 5743 bp long, encoding a protein product of 634 amino acids containing a BTB domain. The protein is highly conserved across different species. Western blot analysis indicates that the KLHL31 protein is abundantly expressed in both embryonic skeletal and heart tissue. In COS-7 cells, KLHL31 proteins are localized to both the nucleus and the cytoplasm. In primary cultures of nascent mouse cardiomyocytes, the majority of endogenous KLHL31 proteins are localized to the cytoplasm. KLHL31 acts as a transcription repressor when fused to GAL4 DNA-binding domain and deletion analysis indicates that the BTB domain is the main region responsible for this repression. Overexpression of KLHL31 in COS-7 cells inhibits the transcriptional activities of both the TPA-response element (TRE) and serum response element (SRE). KLHL31 also significantly reduces JNK activation leading to decreased phosphorylation and protein levels of the JNK target c-Jun in both COS-7 and Hela cells. These results suggest that KLHL31 protein may act as a new transcriptional repressor in MAPK/JNK signaling pathway to regulate cellular functions.

Genetic Studies on the Sea Urchin Embryogenesis and Skeletogenesis (성게의 발생과 뼈대형성의 유전학적 연구)

  • Lee, Youn-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.6 no.4
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    • pp.265-273
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    • 2001
  • The sea urchin has been used as sea food in many countries. This species has also been an important organism of embryological studies for more than a century. In recent years, sea urchin embryos are being used as testing materials for toxicity of pollutants and toxins. Usefulness of sea urchin embryos as experimental models comes from the easiness in obtaining sea urchin samples and a lot of gametes, in rearing embryos in the laboratory, in observing the cellular movement and organ formation during the embryogenesis and in manipulating blastomeres and genetic maferials. The sea urchin in itself is a key organism for the understanding of deuterostome evolution from the protostomes and of indirect development of marine invertebrates which undergo the planktotrophic larval stage. A fertilized sea urchin egg goes through rapid cleavage and becomes a 60 cell embryo 7hr after fertilization. It then develops into a morula, a blastula, a gastrula and finally a pluteus larva approximately 70 hr after fertilization. At the 60 cell stage, the embryo comprises of five territories that express territory-speciflc genes and later form different organs. Micromeres at the vegetal pole ingress into the blastoceol and become the primary mesenchyme cells(PMCs). PMCs express genes involved in skeletogenesis such as SM30, SM37, SM50, PM27, msp130. Among the genes, SM37 and SM50 are considered to be members of a gene family which is characterized by early blastula expression, Glycine-Proline-Glutamine rich repeat structures and spicule matrix forming basic proteins. Genetic studies on the sea urchin embryos help understand the molecular basis of indirect development of marine invertebrates and also of the biomineralization common to the animal kingdom.

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Molecular Characterization of Hanwoo Glucose Transporter 4 Gene (한우 Glucose Transporter 4 유전자의 분자생물학적 해석)

  • Lee, S.M.;Jeong, Y.H.;Kim, H.M.;Park, H.Y.;Yoon, D.H.;Moon, S.J.;Chung, E.R.;Kang, M.J.
    • Journal of Animal Science and Technology
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    • v.47 no.6
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    • pp.1087-1094
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    • 2005
  • The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose transport protein. In the glucose transport protein family, GLUT4 plays a key role in cellular glucose uptake stimulated by insulin in skeletal muscles and adipose tissue in rodents and human. In this studies, we reported the identification, characterization, and expression of Hanwoo GLUT4 gene. The Hanwoo GLUT4 cDNA includes a 1527 bp open reading frame encoding a protein of 509 amino acids. The GLUT4 amino acid sequences of the Hanwoo show strong conservation with the corresponding sequences reported in other species. The highest mRNA expression of GLUT4 was detected in heart and lower expression was detected in rib meat, sirloin, and colon. We confirmed the expression of GLUT4 in the subcutaneous and small intestinal adipose tissue using RT-PCR. To investigate the expression of GLUT4 in the bovine intramuscular adipose differentiation, fibroblast-like cells were isolated from the sirloin of Hanwoo bull aged 12 months by collagenase digestion of minced tissue and cultured with activators of PPAR gamma. We identified that GLUT4 mRNA expression decreased during differentiation of preadipocytes into adipocyte in Korean cattle. These results indicated that function of GLUT4 in bovine adipose tissue was different from that of mouse and human.