• Title/Summary/Keyword: ncRNA

Search Result 162, Processing Time 0.033 seconds

Development of Potato Virus Y Resistant Tobacco Plant by Transformation of the Untranslatable Viral Coat Protein Encoding cDNA (감자 바이러스 Y 비전이성 외피단백질 cDNA의 형질전환에 의한 바이러스 저항성 연초품종 개발)

  • 이청호;이영기;강신웅;박성원;김상석;박은경
    • Journal of the Korean Society of Tobacco Science
    • /
    • v.19 no.2
    • /
    • pp.117-123
    • /
    • 1997
  • Viral coat protein (CP) encoding cDNA with artificial start and stop codons was synthesized by reverse-transcriptase polymerase chain reaction (RT-PCR) from the Korean isolate of potato virus Y-vein nectrosis strain (pVY-VN). To make PVY CP cDNA to untranslatable form, three stop codons were inserted near the start codon by "megaprimer-PCR" method. The untranslatable CP cDNA was subcloned to plant expression vector and transferred to N. tabacum cv. NC82 by Agrobacterium-mediated transformation. Highly resistant plants to PVY infection were screened, based on symptom development after mechanical virus inoculation. By genomic PCR and Southern blot analysis, one or more copies of the untranslatable CP gene were found in all transformants. From northern blot analysis, highly resistant transgenic lines had very low level of CP transcript but susceptible lines had high level, suggesting resistance to PVY infection should be related to RNA-mediated mechanism.mechanism.

  • PDF

Isolation of Coat Protein Gene from Cucumber Mosaic Virus and Its Introduction into Tobacco (오이 모자이크 바이러스 외피 단백질 유전자 분리 및 담배로의 형질전환)

  • 손성한;김경환;김영태;박종석;김주곤;이광웅;황영수
    • Korean Journal of Plant Tissue Culture
    • /
    • v.22 no.3
    • /
    • pp.149-155
    • /
    • 1995
  • The coat protein (CP) gene was cloned from RNA genome of the Cucumber Mosaic Virus strain ABI (CMV-ABI) isolated in Korea. The comparisons of the nucleotide sequence of the cloned CP gene and its deduced amino acid sequences with other CP genes revealed that the CMV-ABI belongs to subgroup I (type I), CMV-ABI developed the typical mosaic symptom in infected plants. Tobacco plants (Samsun and NC82) were transformed by leaf-disc transformation via Agrobacterium, temefaciens LB4404 harboring pVCP, witch CMV-ABI CP gene was inserted into the pBI121, and a number of mature transgenic tobacco plants were developed. Southern and PCR analysis of genomic DNA from the transgenic plants showed that the CP gene was integrated into the genomes of the most of the transgenic plant. Result of the segregation patterns of resistance in T1 seedlings of the plants to kanamycin showed that the transgenic plants containing l,2 and 3 copies of CP gene were50%, 39% and 11% of the total transgenic plants, respectively.

  • PDF

Rifampicin Alleviates Atopic Dermatitis-Like Response in vivo and in vitro

  • Kim, Seung Hyun;Lee, Ki Man;Lee, Geum Seon;Seong, Ju-Won;Kang, Tae Jin
    • Biomolecules & Therapeutics
    • /
    • v.25 no.6
    • /
    • pp.634-640
    • /
    • 2017
  • Atopic dermatitis (AD) is a common inflammatory skin disorder mediated by inflammatory cells, such as macrophages and mast cells. Rifampicin is mainly used for the treatment of tuberculosis. Recently, it was reported that rifampicin has anti-inflammatory and immune-suppressive activities. In this study, we investigated the effect of rifampicin on atopic dermatitis in vivo and in vitro. AD was induced by treatment with 2, 4-dinitrochlorobenzene (DNCB) in NC/Nga mice. A subset of mice was then treated with rifampicin by oral administration. The severity score and scratching behavior were alleviated in the rifampicin-treated group. Serum immunoglobulin E (IgE) and interleukin-4 (IL-4) levels were also ameliorated in mice treated with rifampicin. We next examined whether rifampicin has anti-atopic activity via suppression of mast cell activation. Rifampicin suppressed the release of ${\beta}$-hexosaminidase and histamine from human mast cell (HMC)-1 cultures stimulated with compound 48/80. Treatment with rifampicin also inhibited secretion of inflammatory mediators, such tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$) and prostaglandin $D_2$ ($PGD_2$), in mast cells activated by compound 48/80. The mRNA expression of cyclooxygenase 2 (COX-2) was reduced in the cells treated with rifampicin in a concentration-dependent manner. These results suggest that rifampicin can be used to treat atopic dermatitis.

Taxonomy of Arthrinium minutisporum sp. nov., Pezicula neosporulosa, and Acrocalymma pterocarpi: New Records from Soil in Korea

  • Das, Kallol;Lee, Seung-Yeol;Choi, Hyo-Won;Eom, Ahn-Heum;Cho, Young-Je;Jung, Hee-Young
    • Mycobiology
    • /
    • v.48 no.6
    • /
    • pp.450-463
    • /
    • 2020
  • The strains 17E-042, 17E-039, and NC13-171 belong to Ascomycota and were isolated from soil collected from Sancheong-gun and Yeongam-gun, Korea. The strain 17E-042 produced white mycelial colonies that developed a sienna color with a round margin on potato dextrose agar (PDA), and the reverse side developed a light sienna color. Morphologically, this strain was similar to the strains of Arthrinium phragmites and A. hydei, but the shorter conidial size of the newly identified strain (17E-042) was distinct. The strain 17E-039 produced macroconidia that were pale yellow to orange-brown, elongated-ellipsoid to oblong, round at both ends, primarily straight but sometimes slightly curved, 0-septate, thin-walled, and filled with numerous droplets, having diameters of 20.4-34.3 × 8.0-12.0 ㎛. And the strain NC13-171 formed hyaline to light brown chlamydospores, solitary or in a chain. Multigene phylogenetic analyses were conducted using sequence data obtained from internal transcribed spacer (ITS) regions, 28S rDNA large subunit (LSU), β-tubulin (TUB2), translation elongation factor 1-alpha (TEF1-α), and RNA polymerase II large subunit (RPB2) genes. The results of molecular phylogeny, the detailed descriptions and illustrations of each species strongly support our proposal that these strains from soil in Korea be designated as Arthrinium minutisporum sp. nov. and two new records of Pezicula neosporulosa and Acrocalymma pterocarpi.

The Synergism of Human Lactobacillaceae and Inulin Decrease Hyperglycemia via Regulating the Composition of Gut Microbiota and Metabolic Profiles in db/db Mice

  • Peifan Li;Tong Tong;Yusong Wu;Xin Zhou;Michael Zhang;Jia Liu;Yongxin She;Zuming Li;Yongli Li
    • Journal of Microbiology and Biotechnology
    • /
    • v.33 no.12
    • /
    • pp.1657-1670
    • /
    • 2023
  • This study aimed to evaluate the effects of Limosilactobacillus fermentum and Lactiplantibacillus plantarum isolated from human feces coordinating with inulin on the composition of gut microbiota and metabolic profiles in db/db mice. These supplements were administered to db/db mice for 12 weeks. The results showed that the Lactobacillaceae coordinating with inulin group (LI) exhibited lower fasting blood glucose levels than the model control group (MC). Additionally, LI was found to enhance colon tissue and increase the levels of short-chain fatty acids. 16S rRNA sequencing revealed that the abundance of Corynebacterium and Proteus, which were significantly increased in the MC group compared with NC group, were significantly decreased by the treatment of LI that also restored the key genera of the Lachnospiraceae_NK4A136_group, Lachnoclostridium, Ruminococcus_gnavus_group, Desulfovibrio, and Lachnospiraceae_UCG-006. Untargeted metabolomics analysis showed that lotaustralin, 5-hydroxyindoleacetic acid, and 13(S)-HpODE were increased while L-phenylalanine and L-tryptophan were decreased in the MC group compared with the NC group. However, the intervention of LI reversed the levels of these metabolites in the intestine. Correlation analysis revealed that Lachnoclostridium and Ruminococcus_gnavus_group were negatively correlated with 5-hydroxyindoleacetic acid and 13(S)-HpODE, but positively correlated with L-tryptophan. 13(S)-HpODE was involved in the "linoleic acid metabolism". L-tryptophan and 5-hydroxyindoleacetic acid were involved in "tryptophan metabolism" and "serotonergic synapse". These findings suggest that LI may alleviate type 2 diabetes symptoms by modulating the abundance of Ruminococcus_gnavus_group and Lachnoclostridium to regulate the pathways of "linoleic acid metabolism", "serotonergic synapse", and" tryptophan metabolism". Our results provide new insights into prevention and treatment of type 2 diabetes.

The Effects of Either Chrysin or Moderate Exercise on Inflammasome and Thermogenic Markers in High Fat Fed Mice (고지방식이 동물의 간 조직에서 크리신 투여 또는 중강도 운동이 Inflammasome과 열 발생 유전자발현에 미치는 효과)

  • Lee, Young-Ran;Park, Hee-Geun;Lee, Wang-Lok
    • Journal of Life Science
    • /
    • v.29 no.5
    • /
    • pp.607-613
    • /
    • 2019
  • The purpose of this study was to investigate the effects of either chrysin or exercise on the inflammasome and thermogenic markers in the livers of high-fat fed mice. C57BL/6 mice were randomly assigned to four groups: normal diet control (NC; n=5), high-fat diet control (HC; n=5), high-fat diet with chrysin (Hch; n=5), and high-fat diet with moderate exercise (HME; n=5). The mice were fed a high-fat diet (60% of calories from fat) or normal diet (18% of calories from fat). Chrysin was supplemented orally as 50mg/kg/day dissolved in a 0.1ml solution of dimethyl sulfoxide. The exercised mice ran on a treadmill at 12-20 m/min for 30-60 min/day, 5 times/week, for 16 weeks. After the intervention, the epididymal fat and liver weights were significantly decreased in the HME group compared with HC and Hch groups. The adipocyte size was effectively decreased in the Hch and HME groups compared with the HC group. The inflammasome markers NLRP3, $IL-1{\beta}$, and caspase1 were significantly decreased in the Hch and HME groups compared with the HC group. The thermogenic markers $PGC-1{\alpha}$ and BMP7 were significantly lower in the HC than in the NC group. However, the HME group showed an increase in the thermogenic markers. In conclusion, chrysin and moderate exercise have positive effects on obese metabolic complications induced by high-fat diets by reducing inflammasome genes. However, chrysin supplementation had no effect on thermogenic gene expression. Moderate exercise would therefore seem to be more effective in controlling obesity-induced metabolic deregulation.

Superoxide Dismutase Gene Expression in the Endotoxin-Treated Rat Lung (내독소에 의한 백서 폐장의 Superoxide Dismutase 유전자 발현에 관한 연구)

  • Yoo, Chul-Gyu;Suh, Gee-Young;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
    • /
    • v.41 no.3
    • /
    • pp.215-221
    • /
    • 1994
  • Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

  • PDF

Fermentation and Quality Characteristics of Cheonggukjang Fermented with Bacillus subtilis BC-P1 (Bacillus subtilis BC-P1 균주를 이용하여 제조한 청국장의 발효 및 품질 특성분석)

  • Park, Sung-Yong;Bang, Mi-Ae;Oh, Boung-Jun;Park, Jeong-Hoon;Song, Won-Seob;Choi, Kyung-Min;Choung, Eui-Su;Boo, Hee-Ock;Cho, Seung-Sik
    • Korean Journal of Microbiology
    • /
    • v.49 no.3
    • /
    • pp.262-269
    • /
    • 2013
  • The object of this study was to improve the quality of Cheonggukjang with new starter, Bacillus subtilis BC-P1. Twenty strains were isolated from the commercial cheonggukjang and 1 Bacillus strain (BC-P1) with protease activity was selected. The 16S rRNA gene sequence revealed that the BC-P1 was closely related to B. subtilis with 99% homology. The quality characteristics of chunggukjang fermented with B. subtilis BC-P1, Bacillus nato (PC) and commercial chunggukjang (NC) were investigated. The characteristics of fermentation were determined by protease, lipase, xylanase, chitinase, and fibrinolytic activities, reducing sugar, nutrient composition and amino acid contents of cheonggukjang sample. Cheonggukjang fermented with B. subtilis BC-P1 showed the strongest fibrinolytic, xylanase, and chitinase activities. Reducung sugar contents of Cheonggukjang samples were $30.16{\pm}2.11$ mg/g (NC), $28.56{\pm}1.52$ mg/g (PC), $32.39{\pm}1.87$ mg/g (BC-P1). And their total amino acid contents were 338.99 mg% (NC), 445.19 mg% (PC), 741.35 mg% (BC-P1). These results suggested that B. subtilis BC-P1 was suitable to be used as a starter to enhance the quality and effects of cheonggukjang.

Inhibitory Effects of a Korean Strain Gpf01 Identified as Pseudomonas fluorescens on Cucumber mosaic virus

  • Ipper, Nagesh S.;Kim, Jung-Eun;Koo, Jun-Hak;Hur, Jang-Hyun;Lim, Chun-Keun
    • The Plant Pathology Journal
    • /
    • v.21 no.3
    • /
    • pp.262-269
    • /
    • 2005
  • An antiviral producing bacterial strain was isolated from a ginseng rhizosphere in Kangwon province of Republic of Korea. In order to identify the bacterial strain, microbiological, physiological and biochemical tests were performed, along with RAPD, 16S rRNA, 16S-23S rRNA ITS (intergenic spacer region) and DNA-DNA hybridization analyses. The bacterium was found to be a strain of Pseudomonas fluorescens, which was designated as Gpf01. The strain was grown in Muller-Hinton (MH) broth, and the culture supernatant obtained was filtered through a $0.45{\mu}l$ filter. It was further boiled at $100^{\circ}C$ and tested in two experiments for its ability to control a yellow strain of Cucumber mosaic virus (CMV-Y). In the first experiment, boiled culture filtrate (RCF) was treated on one half of the leaves of Chenopodium amaranticolor followed by CMV- Y inoculation on both halves. In the second experiment, BCF was treated on the lower leaves of Nicotiana tobacum var. Xanthi-nc, with the CMV-Y mechanically inoculated onto the upper untreated leaves. In the first experiment, BCF treatment was able to considerably reduce the number of viral lesion, and in the second experiment, plants treated with BCF showed no visible viral symptoms compared to the Muller-Hinton (MH) media treated controls 15 days post inoculation (dpi), and remained symptomless throughout the study period. Thus, Gpf01, identified as P. fluorescence, was able to produce an antiviral component in the culture filtrate, which was found to be heat stable, non-phytotoxic and effective in local as well as systemic hosts of CMV.

A Novel Strain of Cucumber mosaic virus Isolated from Lilium longiflorum

  • Jung, Hye-Jin;Ueda, Shigenori;Ryu, Ki-Hyun;Lee, Sang-Yong;Choi, Jang-Kyung
    • The Plant Pathology Journal
    • /
    • v.16 no.6
    • /
    • pp.306-311
    • /
    • 2000
  • A new strain of Cucumber mosaic virus (CMV) from easter lily (Lilium longiflorum), Ly2-CMV, was identified and compared to the well-characterized Mf-CMV (subgroupⅠ) and LS-CMV (subgroupⅡ) by host reaction in several indicator plants, dsRNA analysis, serological property, RT-PCR analysis, restriction enzyme profile of the PCR products and nucleotide sequence of coat protein (CP) gene. Remarkable differences in symptoms of Ly2-CMV were found between Mf-CMV or LS-CMV in tobacco plants and Datura stramoinium. Ly2-CMV induced small necrotic ringspots on the inoculated leaves of Nicotiana tabacum cvs. Xanthi nc and Burley 21 and D. stramonium, and failed to infect these species systemically. Of the indicator plants tested, N. benthamiana only reacted with systemic infection by inoculation of Lr2-CMV. In experiments of dsRNA analysis, serology and RT-PCR of CP gene, Ly2-CMV was come within subgroupⅠ CMV. However, restriction enzyme analysis of the PCR products using MspⅠ showed that Ly2-CMV was distinct to Mf-CMV. The CP gene of Ly2-CMV contains 657 nucleotides, and the nucleotide sequence is similar to that of Mf-CMV. There is also a high degree of conservation between their putative gene products in Ly2-CMV and Mf-CMV, with five amino acid changes in the 218 amino acids of the CPs.

  • PDF