• Title/Summary/Keyword: Candidate gene

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Isolation and Characterization of Kimchi Lactic Acid Bacteria Showing Anti-Helicobacter pylori Activity (Helicobacter pylori 억제능 김치 유산균의 분리와 특성 규명)

  • Lee, Youl;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.36 no.2
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    • pp.106-114
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    • 2008
  • One bacterium, which showed strong antagonistic activity against H. pylori KCCM 41756, was isolated from kimchi. The strain NO1 was designated as Lactobacillus plantarum NO1 based on Gram staining, biochemical properties, and 16S rRNA gene sequencing. The culture medium $(2{\sim}4{\mu}g/ml)$ of Lb. plantarum NO1 reduced $(40{\sim}60%)$ the urease activity of H. pylori KCCM 41756. Lb. plantarum NO1 inhibited the binding of H. pylori to human gastric cancer cell line, AGS cells, by more than 33%. Lb. plantarum NO1 exhibited high viability (maintained initial viable cell count of $10^9CFU/ml$) in 0.05 M sodium phosphate buffer (pH 3.0) for 2 h, in artificial gastricjuice for 2 h and in 0.3%, 0.5% oxgall for 24 h. Hemolysis phenomena did not observed when Lb. plantarum NO1 was incubated in the blood agar media. We concluded that Lb. plantarum NO1 can be a good candidate as a probiotic, harboring anti-H. pylori activity.

Biodegradation of Phenanthrene by Transformant Trametes versicolor MrP1 (구름버섯의 형질전환체 Trametes versicolor MrP1에 의한 Phenanthrene의 생분해)

  • Choi, Yun-Seong;Choi, Hyoung-Tae;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.273-278
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    • 2007
  • As a model compound of PAHs (polycyclic aromatic hydrocarbons) phenanthrene has been regarded as a toxic material, mutagen and carcinogen in various animals. Biodegradation conditions of phenanthrene such as pH, temperature, shaking speed, stabilizer and cofactor of degrading enzymes were investigated with Trametes versicolor and its transformant T. versicolor MrP1 in YMG medium, minimal medium and soil microcosm. T. versicolor MrP1 can overexpress mrp gene encoding Mn-repressed peroxidase that is involved in fungal degradation. Biodegradations of phenanthrene by T. versicolor and T. versicolor MrP1 were optimally performed in conditions of weak-acid (pH 6.0), $30^{\circ}C$, shaken culture and medium containing 5 mM veratryl alcohol or tryptophan. In these optimal conditions, biodegradation of phenanthrene by T. versicolor MrP1 is 31% higher than that of wild type strain in a minimal medium for 20 days. Biodegradation of phenanthrene by T. versicolor MrP1 was also higher than that of wild type in soil microcosm. T. versicolor MrP1 can be a excellent candidate for the bioremediation of PAHs contaminated environments.

Role of Tumor Necrosis Factor-Producing Mesenchymal Stem Cells on Apoptosis of Chronic B-lymphocytic Tumor Cells Resistant to Fludarabine-based Chemotherapy

  • Valizadeh, Armita;Ahmadzadeh, Ahmad;Saki, Ghasem;Khodadadi, Ali;Teimoori, Ali
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8533-8539
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    • 2016
  • Background: B-cell chronic lymphocytic leukemia B (B-CLL), the most common type of leukemia, may be caused by apoptosis deficiency in the body. Adipose tissue-derived mesenchymal stem cells (AD-MSCs) as providers of pro-apoptotic molecules such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), can be considered as an effective anti-cancer therapy candidate. Therefore, in this study we assessed the role of tumor necrosis factor-producing mesenchymal stem cells oin apoptosis of B-CLL cells resistant to fludarabine-based chemotherapy. Materials and Methods: In this study, after isolation and culture of AD-MSCs, a lentiviral LeGO-iG2-TRAIL-GFP vector containing a gene producing the ligand pro-apoptotic with plasmid PsPAX2 and PMDG2 virus were transfected into cell-lines to generate T293HEK. Then, T293HEK cell supernatant containing the virus produced after 48 and 72 hours was collected, and these viruses were transduced to reprogram AD-MSCs. Apoptosis rates were separately studied in four groups: group 1, AD-MSCs-TRAIL; group 2, AD-MSCs-GFP; group 3, AD-MSCs; and group 4, CLL. Results: Observed apoptosis rates were: group 1, $42{\pm}1.04%$; group 2, $21{\pm}0.57%$; group 3, $19{\pm}2.6%$; and group 4, % $0.01{\pm}0.01$. The highest rate of apoptosis thus occurred ingroup 1 (transduced TRAIL encoding vector). In this group, the average medium-soluble TRAIL was 72.7pg/m and flow cytometry analysis showed a pro-apoptosis rate of $63{\pm}1.6%$, which was again higher than in other groups. Conclusions: In this study we have shown that tumor necrosis factor (TNF) secreted by AD-MSCs may play an effective role in inducing B-CLL cell apoptosis.

In vivo Biological Function of a Fibrinolytic Enzyme after Oral Adminstration (혈전용해효소의 경구투여에 의한 생체 내 작용)

  • Lee, Young-Hoon;Lee, Sung-Ho;Park, Ki-Hoon;Choi, Young-Ju;Lee, Sang-Won;Kim, Cheol-Ho;Cho, Soo-Jeong;Gal, Sang-Wan
    • KSBB Journal
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    • v.21 no.6 s.101
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    • pp.433-438
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    • 2006
  • A fibrinolytic enzyme gene (BCF-1) was subcloned to the pEB vector which is high expression vector in the Bacillus host. The enzyme was purified by using FPLC after ammonium sulfate precipitation. The enzyme was oral-administrated to the rat and checked the bleeding time, blood clotting time and fibrinolytic effect of the serum. In the bleeding time retardation test, it was longer about 1.7 fold in the feeding rat than without feeding. The serum of rat feeded with the enzyme had the fibrinolytic activity from 1 hour to 3 hours after oral-administration. After 3 hours from feeding, the fibrinolytic activity was decreased gradually. Also blood clotting time after bleeding was longer than that of control rat. The enzyme could be detected at band of 30,000 Da in the blood by western blotting. The enzyme was not harmful to the all internal organs of the rats. Taken together, the enzyme originated from B. subtilis BB-1 can be a candidate to develop the drug for thrombosis, arteriosclerosis and myocardial infarction.

Establishment of High Throughput Screening System Using Human Umbilical Cord-derived Mesenchymal Stem Cells

  • Park, Eu-Gene;Cho, Tae-Jun;Oh, Keun-Hee;Kwon, Soon-Keun;Lee, Dong-Sup;Park, Seung-Bum;Cho, Jae-Jin
    • International Journal of Oral Biology
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    • v.37 no.2
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    • pp.43-50
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    • 2012
  • The use of high throughput screening (HTS) in drug development is principally for the selection new drug candidates or screening of chemical toxicants. This system minimizes the experimental environment and allows for the screening of candidates at the same time. Umbilical cord-derived stem cells have some of the characteristics of fetal stem cell and have several advantages such as the ease with which they can be obtained and lack of ethical issues. To establish a HTS system, optimized conditions that mimic typical cell culture conditions in a minimal space such as 96 well plates are needed for stem cell growth. We have thus established a novel HTS system using human umbilical cord derived-mesenchymal stem cells (hUC-MSCs). To determine the optimal cell number, hUC-MSCs were serially diluted and seeded at 750, 500, 200 and 100 cells per well on 96 well plates. The maintenance efficiencies of these dilutions were compared for 3, 7, 9, and 14 days. The fetal bovine serum (FBS) concentration (20, 10, 5 and 1%) and the cell numbers (750, 500 and 200 cells/well) were compared for 3, 5 and 7 days. In addition, we evaluated the optimal conditions for cell cycle block. These four independent optimization experiments were conducted using an MTT assay. In the results, the optimal conditions for a HTS system using hUC-MSCs were determined to be 300 cell/well cultured for 8 days with 1 or 5% FBS. In addition, we demonstrated that the optimal conditions for a cell cycle block in this culture system are 48 hours in the absence of FBS. In addition, we selected four types of novel small molecule candidates using our HTS system which demonstrates the feasibility if using hUC-MSCs for this type of screen. Moreover, the four candidate compounds can be tested for stem cell research application.

Synergistic efficacy of LBH and αB-crystallin through inhibiting transcriptional activities of p53 and p21

  • Deng, Yun;Li, Yongqing;Fan, Xiongwei;Yuan, Wuzhou;Xie, Huaping;Mo, Xiaoyang;Yan, Yan;Zhou, Junmei;Wang, Yuequn;Ye, Xianli;Wan, Yongqi;Wu, Xiushan
    • BMB Reports
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    • v.43 no.6
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    • pp.432-437
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    • 2010
  • LBH is a transcription factor as a candidate gene for CHD associated with partial trisomy 2p syndrome. To identify potential LBH-interacting partners, a yeast two-hybrid screen using LBH as a bait was performed with a human heart cDNA library. One of the clones identified encodes ${\alpha}B$-crystallin. Co-immunoprecipitation and GST pull-down assays showed that LBH interacts with ${\alpha}B$-crystallin, which is further confirmed by mammalian two-hybrid assays. Co-localization analysis showed that in COS-7 cells, ${\alpha}B$-crystallin that is cytoplasmic alone, accumulates partialy in the nucleus when co-transfected with LBH. Transient transfection assays indicated that overexpression of LBH or ${\alpha}B$-crystallin reduced the transcriptional activities of p53 and p21, respectively, Overexpression of both ${\alpha}B$-crystallin and LBH together resulted in a stronger repression of the transcriptional activities of p21 and p53. These results showed that the interaction of LBH and ${\alpha}B$-crystallin may inhibit synergistically the transcriptional regulation of p53 and p21.

Current biotechnology for the increase of vegetable oil yield in transgenic plants (식물 지방산 생산량의 증진을 위한 생명공학 연구현황)

  • Lee, Kyeong-Ryeol;Choi, Yun-Jung;Kim, Sun-Hee;Roh, Kyung-Hee;Kim, Jong-Bum;Kim, Hyun-Uk
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.241-250
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    • 2011
  • The most part of vegetable oils is accumulated as storage lipid, triacylglycerol (TAG) in seed and used as energy source when seed is germinated. It is also used as essential fatty acids and energy source for human and animal. Recently, vegetable oils have been more and more an important resource because of the increasing demand of vegetable oils for cooking and industrial uses for bio-diesel and industrial feedstock. In order to increase vegetable oils using biotechnology, over-expressing or repressing the regulatory genes involved in the flow of carbon into lipid biosynthesis is critical during seed development. In this review, we described candidate genes may influence oil amount and investigate their potential for oil increase. Genes involved in the regulation from biosynthesis of fatty acids to the accumulation oils in seed can be classified as follows: First, genes play a role for synthesis precursor molecules for TAG. Second, genes participate in fatty acid biosynthesis and TAG assembly. Lastly, genes encodes transcription factors involved in seed maturation and accumulation of seed oil. Because factors/genes determining oil quantity in seed is complex as mentioned, recently regulation of transcription factors is being considered more favorable approach than manipulate multiple genes for increasing oil in transgenic plants. However, it should be figured out the problem that bad agricultural traits induced by the overexpression of transcription factor gene.

Potential Allelic Association of Microsatellite Markers on Bovine Chromosome 5 with Carcass Traits in Hanwoo (Korean cattle) (Microsatellite 의 대립유전자 빈도를 이용한 한우의 경제형질과의 연관성 규명)

  • Oh, Jae-Don;Kong, Hong-Sik;Cho, Byung-Wook;Lee, Mi-Rang;Jeon, Gwang-Joo;Lee, Hak-Kyo
    • Journal of Life Science
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    • v.18 no.9
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    • pp.1225-1229
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    • 2008
  • A total of 10 polymorphic microsatellite markers on bovine chromosome 5 were used for allelic association tests with phenotypic characteristics in Hanwoo. The data analyzed in this study were collected from 326 steers. Chi-square tests were performed to compare the frequencies of individual alleles between the high and the low breeding value groups. The following breeding values were analyzed for QTL effects. The frequency of allele 239 of DIK2828 showed a significant difference between the high and the low breeding value groups in the breeding value of marbling score (MSBV). The allele 279 of BMC1009 was found to show significant differences in allelic distribution for the breeding value of cold carcass weight (CWBV) and the breeding value of backfat thickness (BFBV) and allele 285 showed significant differences in allelic distribution for CWBV, BFBV, and MSBV. The allele 200 of DIK4329 showed significant differences in allelic distributions for the breeding values of longissimus muscle area (LMABV) and BFBV. In this study, we identified the QTL for carcass traits at around 20 (DIK2828), 41 (BMC1009) and 95 (DIK4329) cM in chromosome 5. The results provided a useful reference for further positional candidate gene research and marker-assisted selection for fat metabolism and carcass traits.

Different expression levels of OsPLS1 control leaf senescence period between indica and japonica-type rice

  • Shin, Dongjin;Kim, Tae-Hun;Lee, Ji-Yun;Cho, Jun-Hyeon;Song, You-Chun;Park, Dong-Soo;Oh, Myeong-Gyu
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.98-98
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    • 2017
  • Leaf senescence is the process of aging in plants. Chlorophyll degradation during leaf senescence has the important role translocating nutrients from leaves to storage organs. The functional stay-green with slow leaf yellowing and photosynthesis activity maintenance has been considered one of strategy for increasing crop productivity. Here, we have identified two QTLs on chromosome 9 and 10 for leaf senescence with chlorophyll content of RIL population derived from a cross between Hanareum 2, early leaf senescence Indica-type variety, and Unkwang, delayed leaf senescence Japonica variety. Among these QTLs, we chose qPLS1 QTL on chromosome 9 for further study. qPLS1 was found to explain 14.4% of the total phenotypic variation with 11.2 of LOD score. Through fine-mapping approach, qPLS1 QTL locus was narrowed down to about 25kb in the marker interval between In/del-4-7-9 and In/del-5-9-4. There are 3 genes existed within 25kb of qPLS1 locus: LOC_Os09g36200, LOC_Os09g36210, and LOC_Os09g36220. Among these genes, transcript level of LOC_Os09g36200 was increased during the leaf senescence stage and the expression level of LOC_Os09g36200 in Indica was higher than in Japonica. Finally, we chose LOC_Os09g36200 as candidate gene and renamed it as OsPLS1-In and OsPLS1-Jp from Indica- and Japonica-type rice, respectively. OsPLS1-In and OsPLS1-Jp overexpressing transgenic plants showed both early leaf senescence phenotype. These results indicate that OsPLS1 functions in chlorophyll degradation and the difference of expression level of OsPLS1 cause the difference of leaf senescence between Indica and Japonica in rice.

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A Major Locus for Quantitatively Measured Shank Skin Color Traits in Korean Native Chicken

  • Jin, S.;Lee, J.H.;Seo, D.W.;Cahyadi, M.;Choi, N.R.;Heo, K.N.;Jo, C.;Park, H.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.11
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    • pp.1555-1561
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    • 2016
  • Shank skin color of Korean native chicken (KNC) shows large color variations. It varies from white, yellow, green, bluish or grey to black, whilst in the majority of European breeds the shanks are typically yellow-colored. Three shank skin color-related traits (i.e., lightness [$L^*$], redness [$a^*$], and yellowness [$b^*$]) were measured by a spectrophotometer in 585 progeny from 68 nuclear families in the KNC resource population. We performed genome scan linkage analysis to identify loci that affect quantitatively measured shank skin color traits in KNC. All these birds were genotyped with 167 DNA markers located throughout the 26 autosomes. The SOLAR program was used to conduct multipoint variance-component quantitative trait locus (QTL) analyses. We detected a major QTL that affects $b^*$ value (logarithm of odds [LOD] = 47.5, $p=1.60{\times}10^{-49}$) on GGA24 (GGA for Gallus gallus). At the same location, we also detected a QTL that influences $a^*$ value (LOD = 14.2, $p=6.14{\times}10^{-16}$). Additionally, beta-carotene dioxygenase 2 (BCDO2), the obvious positional candidate gene under the linkage peaks on GGA24, was investigated by the two association tests: i.e., measured genotype association (MGA) and quantitative transmission disequilibrium test (QTDT). Significant associations were detected between BCDO2 g.9367 A>C and $a^*$ ($P_{MGA}=1.69{\times}10^{-28}$; $P_{QTDT}=2.40{\times}10^{-25}$). The strongest associations were between BCDO2 g.9367 A>C and $b^*$ ($P_{MGA}=3.56{\times}10^{-66}$; $P_{QTDT}=1.68{\times}10^{-65}$). However, linkage analyses conditional on the single nucleotide polymorphism indicated that other functional variants should exist. Taken together, we demonstrate for the first time the linkage and association between the BCDO2 locus on GGA24 and quantitatively measured shank skin color traits in KNC.