• Title/Summary/Keyword: Ds-isolates

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Draft genome sequence of lytic bacteriophage SA7 infecting Staphylococcus aureus isolates (Staphylococcus aureus 분리주를 감염시키는 용균 박테리오파지 SA7의 유전체 염기서열 초안)

  • Kim, Youngju;Lee, Gyu Min;Taizhanova, Assiya;Han, Beom Ku;Kim, Hyunil;Ahn, Jeong Keun;Kim, Donghyuk
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.77-78
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    • 2018
  • Staphylococcus aureus is a Gram-positive and a round-shaped bacterium of Firmicutes phylum, and is a common cause of skin infections, respiratory infections, and food poisoning. Bacteriophages infecting S. aureus can be an effective treatment for S. aureus infections. Here, the draft genomic sequence is announced for a lytic bacteriophage SA7 infecting S. aureus isolates. The bacteriophage SA7 was isolated from a sewage water sample near a livestock farm in Chungcheongnam-do, South Korea. SA7 has a genome of 34,730 bp and 34.1% G + C content. The genome has 53 protein-coding genes, 23 of which have predicted functions from BLASTp analysis, leaving the others conserved proteins with unknown function.

Antioxidant and Antifungal Activity of Endophytic Fungi Associated with Agarwood Trees

  • Hidayat, Asep;Turjaman, Maman;Faulina, Sarah Asih;Ridwan, Fadel;Aryanto, Aryanto;Najmulah, Najmulah;Irawadi, Tun Tedja;Iswanto, Apri Heri
    • Journal of the Korean Wood Science and Technology
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    • v.47 no.4
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    • pp.459-471
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    • 2019
  • Several species of Aquilaria and Gyrinops are native to Indonesia and well known as agarwood-producing trees with a high economic value. Their bioactive compounds have a wide spectrum of uses, such as in medicine and cosmetics. These genera have undergone extensive search for novel bioactive compounds. The purpose of this study was to isolate, identify, and characterize the endophytic fungi community associated with Aquilaria malaccensis, A. microcarpa, Gyrinops versteegii, and A. crassna trees and investigate their bioactive properties as antioxidant agents and antagonists. A total of 50 fungi were successfully isolated from different tissues of the four species of agarwood-producing trees. Two isolates exhibited strong antioxidant activity, namely, Apodus oryzae (R2MC3A, $IC_{50}$ 60.92 mg/mL) and Diaporthe sp. (P1DS1[C], $IC_{50}$ 76.65 mg/mL). Two isolates, Pestalotiopsis theae (P3BS3[B]) and Curvularia sp. (P2CD3A), showed >75% antifungal activity against pathogenic Fusarium solani. The results revealed that endophytic fungi associated with the studied agarwood-producing trees had potential antioxidant and antifungal activities for further applications in biotechnology.

Characterization of Cucumver mosaic virus Isolated from Hydrangea macrophylla for. otaksa (Sieb. et Zucc) Wils. (수국에서 분리한 Cucumber mosaic virus의 특성)

  • 방주희;박선정;이금희;최장경;이상용
    • Research in Plant Disease
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    • v.7 no.1
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    • pp.1-7
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    • 2001
  • An isolate of Cucumber mosaic cucumovirus(CMV) was isolated from Hydrangea macrophylla for. otaksa(Sieb. et Zucc. ) Wils. showing mosaic symptoms, and designated as Hm-CMV. Hm-CMV was characterized by the tests of host range, physical properties, serological properties, RNA and coat protein compositions, and reverse transcription and polymerase chain reaction (RT-PCR) analysis. Twelve species in 4 families were used in the host range test of Hm-CMV and could be differentiated from Y-CMV used as a control CMV by the ringspot and line pattern on inoculated leaves of several tobacco plants. Thevirus produced local lesions on inoculated leaves of Chenopodium amarticolor, C. quinoa and Vigna unguiculata. The physical properties of the virus were as follows; thermal inactivation point(TIP) was 60$\^{C}$, dilution end point (DEP) was 10$\^$-3/, and longevity in vitro (LIP) was 3∼4 days. Hm-CMV was serologically identical to Y-CMV. SDS-polyaciylamide gel electrophoresis(SDS-PAGE) showed one major protein band of about 28 kDa. In RNA or dsRNA analysis, Hm-CMV consisted of four RNA or dsRNA species, but satellite RNA was not detected. In RT-PCR using CMV-common primer and CMV subgroup I-specific primer, bothe amplified expected size of about 490 bp and 200 bp DNA fragments from Hm-CMV, respectively. Restriction enzyme analysis of the 490 bp RT-PCR products using EcoR I and Msp I showed that Hm-CMV belonged to CMV subgroup I. However, Hm-CMV could be differentiated from other CMV subgroup I isolates by RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR).

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Characterization of Cucumber mosaic virus Subgroup II Isolated from Paprika (Capsicum annuum var, grossum) in Korea

  • Choi, Gug-Seoun;Kim, Jae-Hyun;Choi, Jang-Kyung
    • The Plant Pathology Journal
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    • v.18 no.1
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    • pp.6-11
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    • 2002
  • An isolate of Cucumber mosaic virus (CMV), PaFMl-CMV causing malformation on the fruit of paprika (Capsicum annuum var, grossum) was characterized based on biological reactions, serological relationships, and partial nucleotide sequence analyses. PaFMl-CMV was distinguishable from other isolates of CMYI Mf-(subgroup I) and LS-CMV (subgroup II), in terms of its reactions to some host plants. Polyclonal antibody against PaFMl-CMV showed homologous antigenic relationship with LS-CMV, however, the antibody formed a spur between PaFMl- and Mf-CMV, In the comparison of molecular size of dsRNAs of PaFMl-CMV with Mf- and LS-CMV, PaFMl-CMV had a slightly smaller RNAl and larger RNA2, RNA3, and RNA4. When the CDNA product of PaFMl-CMV coat protein (CP) gene was digested with some restriction enzymes, the fragment pattern was identical with that of LS-CMV The nucleotide and amino acid sequences of PaFMl-CMV CP gene were 99.5% and 98.6% identical with LS-CMV respectively. The data indicate that PaFMl-CMV belongs to subgroup II of CMV, which is the first report in Korea.

In Vitro Expression and Antibody Preparation of Rice black-streaked dwarf virus Coat Protein Gene (벼검은줄오갈병바이러스 외피단백질 유전자 단백질 발현과 항혈청 제작)

  • Lee, Bong Choon;Cho, Sang-Yun;Bae, Ju Young;Kim, Sang Min;Shin, Dong Bum;Kim, Sun Lim
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.32-37
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    • 2016
  • In this work, major outer capsid protein (P10) encoded by genome segment S10 of Rice black-streaked dwarf virus (RBSDV) was expressed in Escherichia coli. Genomic dsRNA was extracted from RBSDV-miryang isolate infected rice plants. Based on the sequence of S10 (RBSDV-miryang, GenBank JX994211), a pair of S10 specific primers were designed and used to amplify the fragment encoding the N-part of P10. We amplified the partial gene (S10 1-834 nt) of RBSDV P10 (1-278 aa) by RT-PCR. Amplified RBSDV S10 (1-834 nt) was cloned into the expression vector pET32a (+). Recombinant RBSDV S10 (1-834 nt) was expressed in E. coli BL21(DE3) and purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity column. We successfully obtained P10 partial protein of RBSDV and the purified protein was used to immunize rabbits. The resulting polyclonal antiserum specifically recognized RBSDV from infected plant in both Western blotting and enzyme-linked immunosorbent assay. In this study, we provide purified RBSDV P10 (1-278 aa), which would be good material for the serological study of RBSDV-miryang isolates.

Reovirus and Tumor Oncolysis

  • Kim, Man-Bok;Chung, Young-Hwa;Johnston, Randal N.
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.187-192
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    • 2007
  • REOviruses (Respiratory Enteric Orphan viruses) are ubiquitous, non-enveloped viruses containing 10 segments of double-stranded RNA (dsRNA) as their genome. They are common isolates of the respiratory and gastrointestinal tract of humans but are not associated with severe disease and are therefore considered relatively benign. An intriguing characteristic of reovirus is its innate oncolytic potential, which is linked to the transformed state of the cell. When immortalized cells are transfected in vitro with activated oncogenes such as Ras, Sos, v-erbB, or c-myc, they became susceptible to reovirus infection and subsequent cellular lysis, indicating that oncogene signaling pathways are exploited by reovirus. This observation has led to the use of the virus in clinical trials as an anti-cancer agent against oncogenic tumors. In addition to the exploitation of oncogene signaling, reovirus may further utilize host immune responses to enhance its antitumor activity in vivo due to its innate interferon induction ability. Reovirus is, however, not entirely benign to immunocompromised animal models. Reovirus causes so-called "black feet syndrome" in immunodeficient mice and can also harm neonatal animals. Because cancer patients often undergo immunosuppression due to heavy chemo/radiation-treatments or advanced tumor progression, this pathogenic response may be a hurdle in virus-based anticancer therapies. However, a genetically attenuated reovirus variant derived from persistent reovirus infection of cells in vitro is able to exert potent anti-tumor activity with significantly reduced viral pathogenesis in immunocompromised animals. Importantly, in this instance the attenuated, reovirus maintains its oncolytic potential while significantly reducing viral pathogenesis in vivo.

Characterization of an Isolate of Cucumber mosaic virus Isolated from Canna generalis Bailey (칸나에서 분리한 Cucumber mosaic virus의 특성)

  • Jeon, Yong-Woon;Hong, Jin-Sung;Lee, Sang-Yong;Ryu, Ki-Hyun;Choi, Jang-Kyung
    • Research in Plant Disease
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    • v.12 no.3
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    • pp.298-302
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    • 2006
  • An isolate of Cucumber mosaic virus(CMV), called as Can-CMV, was originally isolated from Canna generalis showing typical streak mosaic foliar symptoms, and its properties were investigated in this study. Whereas all known isolates of CMV could induce symptoms on their systemic hosts(four kinds of Nicotiana spp and a zucchini squash), Can-CMV induced no symptoms on its systemic hosts tested. Replication and movement of the virus on upper leaves as well as inoculated leaves-were confirmed by RT-PCR suggesting that Can-CMV could only infect systemically on N. benthamiana and N. glutinosa. Size of local lesions on the Can-CMV-inoculated leaves of Chenopodium amaranticolor was much smaller than that of Fny-CMV. Whereas Fny-CMV and LS-CMV could induce distinct necrotic local lesions on Vigna unguiculata 2 to 3 days postinoculation(dpi), chlorotic spots symptom was expressed by Can-CMV 4 to 5 dpi. Virus-specific 4 kinds of dsRNAs were isolated from leaves of N. benthamiana infected with Can-CMV, and these dsRNAs corresponded to the viral genomic RNAs and subgenomic RNAs and their patterns were indistinguishable to those of Fny-CMV and LS-CMV. By restriction mapping analysis of 950 bp of RT-PCR amplified products of coat protein gene of the virus as well as by serological analysis of gel diffusion test, Can-CMV belongs to a typical member of CMV subgroup IA. These results suggest that the Can-CMV isolated from C. generalis possesses unique pathological properties to understand further insight into the various interactions between virus and host.

Experimental Pathogenesis of Pullorum Disease with the Local Isolate of Salmonella enterica serovar. enterica subspecies Pullorum in Pullets in Bangladesh

  • Haider, M.G.;Chowdhury, E.H.;Khan, M.A.H.N.A.;Hossain, M.T.;Rahman, M.S.;Song, H.J.;Hossain, M.M.
    • Korean Journal of Poultry Science
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    • v.35 no.4
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    • pp.341-350
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    • 2009
  • The research work was carried out to study the pathogenesis covering the clinical signs, gross and histopathological lesions in different organs, and reisolation and identification of the organisms after experimental infection with the local isolate of Salmonella enterica serovar. enterica subspecies (S.) Pullorum at different time interval of the experiment during the period February 2006 to December 2006. One hundred pullets (seronegative to S. Pullorum of 12 weeks age were purchased and divided into 5 (A, B, C, D and E) groups and each group consisted of 20 birds. Four groups (A, B, C and D) were infected orally with a dose of $10^6\;CFU$, $10^7\;CFU$, $2{\times}10^7\;CFU$, $10^8\;CFU$ of S. Pullorum, respectively, and one group (E) was treated as uninfected control. The used methods were necropsy and histopathology, culture of bacteria, staining and biochemical test of Salmonella. Five birds from each group were randomly selected and sacrificed $1^{st}$ week, $2^{nd}$, $3^{rd}$ and $4^{th}$ weeks of post infection (PI). From all the groups, the bacteriological samples (crop, liver, lung, heart, spleen, bile duodenum, ceca and blood) were collected with pre enriched in buffered peptone water in sterile poly bags. Liver, lungs, heart, spleen, intestine, etc. were collected in 10% buffered-formalin for histopathological examination. No clinical signs, gross and histopathological lesions were found in control group and no S. Pullorum was reisolated. Clinical sign of experimentally infected with S. Pullorum in pullets were loss of appetite (100%), slight depression (75%), ruffled feathers (85%), diarrhea (60%) and loss of weight (100%) in chickens. The feed intake and body weight at different weeks after PI differed significantly (p<0.01) among the groups. Grossly, the highest recorded lesion was button-like ulcer in the ceca (80%) and the lowest was white nodules in lungs (1.25%). S. Pullorum were reisolated from crop (91.25%), liver (91.25%), lung (83.75%), heart (71.25%), spleen (87.75%), bile (33.25%), duodenum (92.50%), ceca (97.50%) and from different group of infection (61.25%). The highest microscopic findings were intestinal and cecal mucosa and submucosa exhibited infiltration of mononuclear cells and congestion (96.25%), and the lowest finding was nodule formation in the lungs (3.75%). The pattern of the disease production by local isolate of S. Pullorum in Bangladesh is almost similar with other isolates in different countries.