• Title/Summary/Keyword: Molecular Biology

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Isolation of Agrobacterium sp. BE516 from the Root of Miscanthus sacchariflorus and Its Plant Growth Promoting Activity (물억새 뿌리로부터 Agrobacterium sp. BE516 균주의 분리 및 식물생육촉진활성)

  • Kang, Hye-Young;Park, Dong-Jin;Lee, Jae-Chan;Kwon, Mi-Kyung;Kim, Seung-Bum;Kim, Chang-Jin
    • Journal of Applied Biological Chemistry
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    • v.55 no.2
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    • pp.129-133
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    • 2012
  • To exploit plant growth promoting bacteria in the roots of Miscanthus sacchariflorus, a biomass energy crop, total 64 bacteria were isolated. For the investigation of plant growth promoting effects from the isolated bacteria, production of indole acetic acid (IAA) and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activities were tested and other cultural conditions were examined. As results, 8 isolates showed plant growth promoting effects on the M. sacchariflorus and an isolate designated Agrobacterium sp. BE516 has the highest activity by enhancing the shoot elongation over 2-fold than the control. Agrobacterium sp. BE516 produced 64 ${\mu}g$ IAA per mL and showed ACC deaminase activity which is involved in the resistance to environmental stress such as high salt and drought. It could grow at low temperature in the range from 4 to $15^{\circ}C$, at pH 4.0 and at 4% NaCl. These results indicate that the Agrobacterium sp. BE516 can be useful as a bio-fertilizer for M. sacchariflorus under the stressed conditions.

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.

The Relationship between Water-Bloom and Distribution of Microorganisms That Inhibit the Growth of Cyanobacterium (Anabaena cylindrica) (수화와 시안세균(Anabaena cylindrica) 생장 억제 미생물 분포도의 상관관계)

  • Kim, Chul-Ho;Lee, Jung-Ho;Choi, Yong-Keel
    • Korean Journal of Microbiology
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    • v.34 no.4
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    • pp.188-193
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    • 1998
  • The authors examined the variations of environmental factors, the distributions of cyanobacteria, heterotrophic bacteria, and microorganisms that inhibit the growth of Anabaena cylindrica according to development and extinction of cyanobacterial bloom at a site in Daechung Dam reservoir. And certified the relationship between each other. Water temperature variated in a typical pattern. pH and concentrations of dissolved oxygen and chlorophylla was high in bloom period, and lowered with the decline of bloom. Phosphorus played as a growth-limiting factor at this study site. Total nitrogen concentration increased during blooming period, which indicated that nitrogen has been fixed by aquatic organisms such as cyanobacteria. Cyanobacteria distributed from June 17, and such cyanobacterial species as Anabaena spp., Aphanizomenon spp., Microcystis spp., Oscillatoria spp. and Phormidium spp. was detected during study period. Anabaena spp. distributed relatively highly distributed from July 23 to September 22, and disappeared completely at September 29. Heterotrophic bacterial and cyanobacterial populations varied inverse-proportionally. There was a relevancy between the variations of Anabaena spp., heterotrophic bacteria, and microorganisms that inhibit the growth of Anabaena cylindrica. Microorganisms that inhibit the growth of Anabaena cylindrica distributed from early growth phase of Anabaena spp. population to immediately after the extinction of Anabaena spp. With the population of Anabaena cylindrica growth-inhibiting microorganisms decreasing, increases of heterotrophic bacterial population followed it. Thease results indicate that microorganisms have a part in the extinction of cyanobacterial bloom, especially at its destroying period.

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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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Baicalein Inhibits the Migration and Invasion of B16F10 Mouse Melanoma Cells through Inactivation of the PI3K/Akt Signaling Pathway

  • Choi, Eun-Ok;Cho, Eun-Ju;Jeong, Jin-Woo;Park, Cheol;Hong, Su-Hyun;Hwang, Hye-Jin;Moon, Sung-Kwon;Son, Chang Gue;Kim, Wun-Jae;Choi, Yung Hyun
    • Biomolecules & Therapeutics
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    • v.25 no.2
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    • pp.213-221
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    • 2017
  • Baicalein, a natural flavonoid obtained from the rhizome of Scutellaria baicalensis Georgi, has been reported to have anticancer activities in several human cancer cell lines. However, its antimetastatic effects and associated mechanisms in melanoma cells have not been extensively studied. The current study examined the effects of baicalein on cell motility and anti-invasive activity using mouse melanoma B16F10 cells. Within the noncytotoxic concentration range, baicalein significantly inhibited the cell motility and invasiveness of B16F10 cells in a concentration-dependent manner. Baicalein also reduced the activity and expression of matrix metalloproteinase (MMP)-2 and -9; however, the levels of tissue inhibitor of metalloproteinase-1 and -2 were concomitantly increased. The inhibitory effects of baicalein on cell motility and invasiveness were found to be associated with its tightening of tight junction (TJ), which was demonstrated by an increase in transepithelial electrical resistance and downregulation of the claudin family of proteins. Additionally, treatment with baicalein markedly reduced the expression levels of lipopolysaccharide-induced phosphorylated Akt and the invasive activity in B16F10 cells. Taken together, these results suggest that baicalein inhibits B16F10 melanoma cell migration and invasion by reducing the expression of MMPs and tightening TJ through the suppression of claudin expression, possibly in association with a suppression of the phosphoinositide 3-kinase/Akt signaling pathway.

Construction of Gene-Specific Primers for Various Antioxidant Isoenzyme Genes and Their Expressions in Rice (Oryza sativa L.) Seedlings Obtained from Gamma-irradiated Seeds

  • Kim, Jin-Hon;Chung, Byung-Yeoup;Kim, Jae-Sung;Wi, Seung-Gon;Yang, Dae-Hwa;Lee, Choon-Hwan;Lee, Myung-Chul
    • Journal of Photoscience
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    • v.11 no.3
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    • pp.115-120
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    • 2004
  • For the expression study of antioxidant isoenzyme genes in rice (Oryza sativa L.) plants, extensive searches for genes of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) isoforms were performed through the GenBank database. The genes for two cytosolic and one plastidic CuZn-SOD, one Fe-SOD, two Mn-SOD, two cytosolic and two chloroplastic (stromal and thylakoid) APX, and three CAT isoforms were available in japonica-type rice. These isoforms were named as cCuZn-SOD1, cCuZn-SOD2, pCuZn-SOD, Fe-SOD, Mn-SOD1, Mn-SOD2, cAPXa, cAPXb, Chl_sAPX, Chl_tAPX, CATa, CATb, and CATc, respectively. Since they shared a high degree of homology in the nucleotide and amino acid sequences, the gene-specific primers for the genes were designed directly from their full-length cDNAs found in the database except for the CATa gene. These primers were used in the RT-PCR analysis to investigate the differential expression of antioxidant isoenzyme genes in rice plants from the seeds irradiated with low doses (2, 4, 8, and 16 Gy) of gamma-radiation. The gammairradiation slightly increased the transcripts of pCuZn-SOD, while those of Fe-SOD, cAPXb, and CATb decreased. However, no substantial differences were observed in the expression of all the isoenzyme genes between the control and irradiated groups. In this study, gene specific primers for thirteen SOD, APX and CAT isoenzymes were constructed from the full-length cDNAs. The results of RT-PCR analysis obtained by using these primers suggests that the expression levels of SOD, APX, and CAT isoenzyme genes in rice seedlings were hardly affected by gamma-irradiation at the seed stage.

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Induction of Apoptosis by Methanol Extract of Gloiopeltis furcata in Human Leukemia Cell Line U937 (인체백혈병세포의 증식에 미치는 불등가사리 메탄올 추출물의 영향)

  • Choi, Woo Young;Park, Cheol;Kim, Gi Young;Lee, Won Ho;Bae, Song-Ja;Choi, Yung Hyun
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.2
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    • pp.76-83
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    • 2006
  • Epidemiological studies have indicated that the ubiquitous consumption of seaweeds is a protective factor against some types of cancer. Previous results showed that the administration of seaweed powder or extract reduced the incidence rate of chemically induced tumorigenesis using in vivo animal model. Recently, we reported that the extracts of Gloiopeltis furcata, a kind of Korean edible seaweed, caused he cell growth inhibition of various human cancer cell lines, among them methanol extract exhibited a relatively strong antiproliferative activity. However, the molecular mechanisms of this seaweed in malignant cells have been poorly studied until now. To elucidate this problem, we investigated the effects of methanol extract of G. furcata (MEGF) on the growth inhibition in several human cancer cell lines, and further we analyzed the effects of this extract were tested on the activity of apoptosis induction in human leukemic cells. The results demonstrated that MEGF treatment resulted in the morphological changes and the growth inhibition in a dose-dependent manner. Furthermore, MEGF potently suppresses the growth of human leukemic U937 cells by induction of apoptosis, which was associated with induction of cyclin-dependent kinase inhibitor p21(WAF1/CIP1) in a tumor suppressor p53-independent fashion and up-regulation of Fas/FasL system. Further studies will be needed to identify the active compounds that confer the anticancer activity of MEGF. Once such compounds are identified, the mechanisms by which they exert their effects can begin to be characterized.

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Gene Promoter Variation of Phosphoglycerate Kinase, a Glucose Metabolism Enzyme, is a Biomarker for Selection of Disease-resistant Sea Squirt, Halocynthia Roretzi (당 생합성 효소 PGK 유전자 프로모터 변이와 물렁증 저항성 멍게의 선별)

  • Cho, Hyun Kook;Hur, Young Baek;Cheong, Jae Hun
    • Journal of Life Science
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    • v.23 no.2
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    • pp.190-196
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    • 2013
  • The sea squirt, Halocynthia roretzi, has experienced mass mortality due to softness syndrome. The identification of disease-induced genes can provide insights into the development of this syndrome. To identify the genes, we performed differentially expressed gene (DEG) analysis. The expression of the phosphoglycerate kinase (HrPGK) gene was significantly decreased in diseased sea squirts compared to normal ones. We confirmed the result of the DEG analysis through RT-PCR and real-time PCR. In addition, we detected single nucleotide polymorphisms at position -106 (A/T) and -254 (G/T) in the HrPGK gene promoter by genotyping analysis. At the -106 site of the HrPGK gene, the frequency of the AA allele in disease-resistant sea squirts was about two-fold higher than that of sensitive ones, and the frequency of the TT allele in the disease-resistant sea squirts was about six-fold lower. At the -254 site of the HrPGK gene, the frequency of the GT and the GG allele was approximately two-fold higher and two-fold lower, respectively, in the disease-resistant sea squirts compared to the disease-sensitive ones. Analysis of the relationship between the genotypic variation at the -106/-254 promoter and the expression of HrPGK mRNA showed that HrPGK mRNA expression was higher in the -106/-254 AA/GT genotype samples than in the -106/254 TT/GG genotype ones. These results show that sea squirts harboring the AA/GT genotype may have more resistance to mortality than the sea squirts with other genotypes.

Establishment and Characterization of Three Immortal Bovine Muscular Epithelial Cell Lines

  • Jin, Xun;Lee, Joong-Seob;Kwak, Sungwook;Lee, Soo-Yeon;Jung, Ji-Eun;Kim, Tae-Kyung;Xu, Chenxiong;Hong, Zhongshan;Li, Zhehu;Kim, Sun-Myung;Pian, Xumin;Lee, Dong-Hee;Yoon, Jong-Taek;You, Seungkwon;Choi, Yun-Jaie;Kim, Hyunggee
    • Molecules and Cells
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    • v.21 no.1
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    • pp.29-33
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    • 2006
  • We have established three immortal bovine muscular epithelial (BME) cell lines, one spontaneously immortalized (BMES), the second SV40LT-mediated (BMEV) and the third hTERT-mediated (BMET). The morphology of the three immortal cell lines was similar to that of early passage primary BME cells. Each of the immortal cell lines made cytokeratin, a typical epithelial marker. BMET grew faster than the other immortal lines and the BME cells, in 10% FBS-DMEM medium, whereas neither the primary cells nor the three immortal cell lines grew in 0.5% FBS-DMEM. The primary BME cells and the immortal cell lines, with the exception of BMES, made increasing amounts of p53 protein when treated with doxorubicin, a DNA damaging agent. On the other hand, almost half of the cells in populations of the three immortal cell lines may lack $p16^{INK4a}$ regulatory function, compared to primary BME cells that were growth arrested by enforced expression of $p16^{INK4a}$. In soft-agar assays, the primary cells and immortal cell lines proved to be less transformed in phenotype than HeLa cells. The three immortal epithelial-type cell lines reported here are the first cell lines established from muscle tissue of bovine or other species.

Sphingosine 1-Phosphate-induced Signal Transduction in Cat Esophagus Smooth Muscle Cells

  • Song, Hyun Ju;Choi, Tai Sik;Chung, Fa Yong;Park, Sun Young;Ryu, Jung Soo;Woo, Jae Gwang;Min, Young Sil;Shin, Chang Yell;Sohn, Uy Dong
    • Molecules and Cells
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    • v.21 no.1
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    • pp.42-51
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    • 2006
  • We investigated the mechanism of contraction induced by S1P in esophageal smooth muscle cells. Western blot analysis demonstrated that $S1P_1$, $S1P_2$, $S1P_3$, and $S1P_5$ receptors existed in the cat esophagus. Only penetration of EDG-5 ($S1P_2$) antibody into permeabilized cells inhibited S1P-induced contraction. Pertussis toxin (PTX) also inhibited contraction, suggesting that it was mediated by $S1P_2$ receptors coupled to a PTXsensitive $G_i$ protein. Specific antibodies to $G_{i2}$, $G_q$ and $G_{\beta}$ inhibited contraction, implying that the S1P-induced contraction depends on PTX-insensitive $G_q$ and $G_{\beta}$ dimers as well as the PTX-sensitive $G_{i2}$. Contraction was not affected by the phospholipase $A_2$ inhibitor DEDA, or the PLD inhibitor ${\rho}$-chloromercuribenzoate, but it was abolished by the PLC inhibitor U73122. Incubation of permeabilized cells with $PLC{\beta}3$ antibody also inhibited contraction. Contraction involved the activation of a PKC pathway since it was affected by GF109203X and chelerythrine. Since $PKC{\varepsilon}$ antibody inhibited contraction, $PKC{\varepsilon}$ may be required. Preincubation of the muscle cells with the MEK inhibitor PD98059 blocked S1P-induced contraction, but the p38 MAP kinase inhibitor SB202190 did not. In addition, co-treatment of cells with GF 109203X and PD98059 did not have a synergistic effect, suggesting that these two kinases are involved in the same signaling pathway. Our data suggest that S1P-induced contraction in esophageal smooth muscle cells is mediated by $S1P_2$ receptors coupled to PTX-sensitive $G_{i2}$ proteins, and PTX-insensitive $G_q$ and $G_{\beta}$ proteins, and that the resulting activation of the $PLC{\beta}3$ and $PKC{\varepsilon}$ pathway leads to activation of a p44/p42 MAPK pathway.