• Title/Summary/Keyword: highly resistant strain

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BmNPV Infection Enhances Ubiquitin-conjugating Enzyme E2 Expression in the Midgut of BmNPV Susceptible Silkworm Strain

  • Gao, Lu;Chen, Keping;Yao, Qin;Chen, Huiqing
    • International Journal of Industrial Entomology and Biomaterials
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    • v.13 no.1
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    • pp.31-35
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    • 2006
  • The ubiquitin conjugating enzyme 2 (E2) is core component of ubiquitin proteasome pathway (UPP) which represents a selective mechanism for intracellular proteolysis in eukaryotic cells. The E2 has been implicated in the intracellular transfer of ubiquitin to target protein. We show here the involvement of E2 in antiviral immune of Bombyx mori to Bombyx mori nuclear polyhedrosis virus (BmNPV). In this study, mRNA fluorescent differential display PCR (FDD-PCR) was performed with BmNPV highly resistant silkworm strain NB and susceptible silkworm strain 306. At 24 h post BmNPV infection, FDD-PCR with the arbitrary primer AP34 showed that one cDNA band was down-regulated in the midgut of resistant strain, but highly expressed in susceptible strain. The deduced amino acid sequence of this cDNA clone share 99% identity with the recently published B. mori ubiquitin conjugating enzyme E2 (Genbank NO: DQ311351). Fluorescent quantitative PCR corroborated down regulation of E2 in resistant strain. We there conclude that BmNPV infection evokes strong response of susceptible strain including activation of UPP. BmNPV may evolve escape mechanisms that manipulate the UPP in order to persist in the infected host. In addition, the identification of down-regulation of E2 in resistant strain, as well as structure data, are essential to understanding how UPP operates in silkworm antiviral immune to BmNPV disease.

Resistance to Potato Virus Y Conferred by PVY Replicase Gene Sequence in Transgenic Burley Tobacco (감자바이러스 Y 복제 유전자로 형질전환된 버어리종 연초의 PVY에 대한 저항성 특성)

  • Young Ho Kim;Eun Kyung Park;Soon Yong Chae;Sang Seock Kim;Kyung-Hee Paek;Hye Sun Cho
    • Journal of the Korean Society of Tobacco Science
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    • v.20 no.1
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    • pp.50-56
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    • 1998
  • The complementary DNA (cDNA) of potato virus Y- vein necrosis strain (PVY-VN) replicase gene (Nlb) was transformed into tobacco (Nicotiana tabacum cv. Burley 21) plants. Out of 25 putative transformants regenerated, 3 were resistant to PVY-VN, one highly resistant plant with no symptom until seed harvest time and the other two with mild chlorotic spot symptoms at late stages after infection. No symptom was observed in the highly resistant plant, while mild vein necrotic symptoms were developed on suckers of the moderately resistant plants after seed harvest time, In the first generation (T1) via self fertilization, resistance to susceptibility frequency in transgenic plants from the highly resistant transformant was about 3 : 1, while it was lowered much (about 1:2 and 1:19) in T1 of the moderately resistant transformants. In the second generation (T2) of the highly resistant plant, resistance frequencies were similar to T1, but resistance levels varied greatly and appeared to be decreased. Key words : potato virus Y, viral replicate gene, transgenic tobacco plants, resistance.

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Design of Metal Cored Wire for Erosion Resistant Overlay Welding

  • Kim, Jun-Ki;Kim, In-Ju;Kim, Ki-Nam;Kim, Ji-Hui;Kim, Seon-Jin
    • Proceedings of the KWS Conference
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    • 2009.11a
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    • pp.202-204
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    • 2009
  • Erosion is a common failure mode of materials frequently encountered in plant and power industry. Although the erosion resistance of Fe-base alloy has been inferior to the other expensive materials, it is expected that the strain-induced martensitic transformation can impart high erosion resistance to Fe-base alloy. The key technology to develop Fe-base metal cored welding wire for erosion resistant overlay welding may include the strain-induced metallurgy for hardening rate control and the welding flux metallurgy for dilution control. Sophisticated studies showed that the strain-induced martensitic transformation behavior was related to the critical strain energy which was dependent on the alloy composition. Dilution and bead shape of overlay weld were proved to be affected by metal transfer mode during gas tungsten arc welding and elements in welding fluxes. It was considered that the highly erosion resistant Fe-base overlay weld could be achieved by precise control of alloy composition to have proper level of critical strain energy for energy absorption and welding flux formulation to have small amount of deoxidizing metallic elements for dilution.

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The Purfication and Characterization of Macrolide-Phosphotransferase K of Escherichia coli 209K Highly Resistant to Erythromycin (에리스로마이신 고도내성 대장균 209K 유래 마크로라이드-포스포트란스페라제 K의 정제 및 특성)

  • Kim, Sook-Kyung;Oh, Tae-Gwon;Baek, Moon-Chang;Hong, Jong-Soo;Kim, Byong-Kak;Choi, Eung-Chil
    • YAKHAK HOEJI
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    • v.41 no.3
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    • pp.359-364
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    • 1997
  • Resistance gene mphK was cloned from Escherichia coli 209K strain which is highly resistant to erythromycin (EM). By using the cloned plasmid pGE64, E. coli NM522 was transformed. The comparison of macrolide-phosphotransferase K [MPH(K)] activity between E. coli 209K and E. coli NM522(pGE64) showed that the total enzyme activity of MN522(pGE64) was fifty-fole higher than that of 209K. To identify characteristics of MPH(K) more precisely. MPH(K) was isolated and purified from the NM522 (pGE64). The final purification f MPH(K) through several stages of purification process was 89 fole and the overall recovery was 11%. This enzyme was monomer with the molecular weight of 34 kDa and its isoelectric point (pI) was 5.0. The optimal pH and temperature for activity were 8.0 and $40^{\circ}C$, respectively.

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Inhibition Effect of Germ-resistant Sponge on Microbial Growth in Kitchen Hygiene (주방위생에서 항균수세미의 효과)

  • 이용욱;나승식;조성범;정지연;박성기
    • Journal of Environmental Health Sciences
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    • v.22 no.4
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    • pp.109-121
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    • 1996
  • It was intended to investigate the effect of the microbiological kitchen hygiene such as dishclothes and scrubbers. The 8indicator organisms (standard plate counts, coliform, heterotroph, enterococcus, staphylococcus, heat-stable bacteria, psychrotroph, Pseudomonas aeruginosa) were detected highly in dishwaters, dishcloth and scrubber. Coliform and Staphylococcus aureus were appeared on dishcloth dominantly than the scrubber, and the scrubbers were intruded by hetrotrophs and psychrotrophs numerously than dishclothes. The germ-resistant sponge inhibited the growth of the most of test strain, and appeared the about 100% reduction rate after 24 hr, but did not affect Pseudomonas aeruginosa and P. fragi so typically after 24 hr. The anti-microorganism durability of germ-resistant sponge, treated with food soil, was maintained by 10 days, the early stage strain density was founded in 20 days, and the strains grew after 30 days.

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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
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    • v.19 no.2
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    • pp.117-123
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    • 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.

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Resistance Characteristics of Flue-cured Tobacco Plants Transformed with CDNA of Potato Virus Y Replicase Gene (감자 바이러스 Y 복제유전자 cDNA로 형질전환된 황색종 담배의 저항성 특성)

  • 박은경;백경희;유진삼;조혜선;강신웅;김영호
    • Journal of the Korean Society of Tobacco Science
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    • v.19 no.1
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    • pp.11-17
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    • 1997
  • A flue-cured tobacco variety (Nicotiana tabacum cv. Wisconsin) was used for Plant transformation with the complementary DNA (cDNA) of potato virus Y-necrosis strain (PVY-VN) replicase gone (Nb) which was synthesized through reverse-transcription Primed with oligo(dT) and Polymerization using RNase H-digested template. The cDNA was cloned into Plant expression vector Plasmid (PMBP2), and introduced into tobacco plants by co-culturing tobacco leaf disks with Agrobacterium tumefaciens LBA4404 containing the plasmid before Plant regeneration. Eight Plants, in which the inserted cDNA fragment was detected by Polymerase chain reaction (PCR), out of 70 putative transformants inserted with sense-oriented Mb cDNA showed no symptom at 3 weeks after inoculation, while the other 62 plants, and all plants with vector gone only and antisense-oriented NIb cDNA had susceptible vein-necrosis symptoms. However, only 2 of the 8 resistant plants were highly resistant, which remained symptomless up to 10 weeks after inoculation. Among the first progenies (T1) from self-fertilized seeds of the two resistant transgenic plants, less than 10 % of 71 plants appeared highly resistant (with no symptom), 70% moderately resistant (with mild symptoms on 1 - 2 leaves), and about 20% susceptible (with susceptible symptoms on 3 or more leaves) at 3 weeks after inoculation. These results suggest that the PVY resistance was inherited in the 71 generation. Key words : potato virus Y. viral replicase gene, transgenic tobacco Plants, resistance.

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Antibiotic Resistance of Bacterial Isolates from Nasal Discharges of Dogs with Respiratory Diseases (개의 비루에서 분리한 원인균의 항생제 내성)

  • 김문선;정종태;강태영;윤영민;이주명;이두식;손원근
    • Journal of Veterinary Clinics
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    • v.21 no.2
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    • pp.133-139
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    • 2004
  • Bacterial pathogens were isolated from 36 dogs with respiratory signs, that were submitted to Veterinary Clinics in Jeju, including Veterinary Medical Teaching Hospital in Cheju National University. Of 36 isolates, 16 (44.4%) bacterial pathogens were Gram-positive and 20 (55.6%) were Gram-negative. Gram-positive bacteria identified with API Staph were 12 S. intermedius (33.3%), 2 S. aureus (5.6%), 1 S. haemolyticum (2.8%), and 1 S. xylosus (2.8%). Gram-negative organisms identified with API 20E or API NE included 8 Bordetella bronchiseptica (22.2%), 6 Escherichia coli (16.7%), 4 Pasteurella spp. (11.1%), 1 Enterobacter intermedius (2.8%), and 1 Oligella ureolytica (2.8%). Both Staphylococcus spp. isolates and Gram-negative pathogens were resistant to one or more antibiotics, including ampicillin (AM), amoxicillin/clavulanic acid (AMC), chloramphenicol (C), cefazolin (CZ), erythromycin (E), gentamicin (GM), kanamycin (K), lincomycin (L), oxacillin (OX), trimethoprim/sulfamethoxazole (SXT), and tetracycline (TE). All Staphylococcus spp. were susceptible to AMC, OX and VA, while many isolates were highly resistant to L (87.5%), E (68.8%), P (62.5%), and AM (56.3%). Antibiotic-resistant patterns of staphylococcal isolates were shown ranges from single to 9-resistant patterns. Resistant rates to antibiotics of Gram-negative bacteria were usually higher than those of Staphylococcus spp. in this study. Most Gram-negative bacteria were highly resistant to L (90.0%), AM (85.0%), E (85.0%), P (85.0%), OX (80.0%), and CZ (75.0%). B. bronchiseptica isolates showed 5 to 8 antibiotics-resistant patterns and Pasteurella spp., 2 to 8-resistant patterns. In particular, all 6 E. coli isolates were resistant to more than 9 different kinds of antibiotics, including one strain resistant to all antibiotics tested.

Isolation and Biological Characterization of Barley mild mosaic virus(BaMMV) Mild and Severe Strains in Korea

  • Jonson, Gilda;Park, Jong-Chul;Noh, Tae-Hwan;Kim, Mi-Jung;Hyun, Jong-Nae;Kim, Jung-Gon
    • The Plant Pathology Journal
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    • v.22 no.4
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    • pp.329-333
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    • 2006
  • Two distinct and stable isolates of Barley mild mosaic virus(BaMMV) designated as Naju82-S(severe) and Naju82-M(mild) were obtained. These two isolates differed in their symptomatology, virus transmission characteristics and cultivar specificity at various temperature. Thus, these isolates were referred to as strains in this study. BaMMV Naju-S strain showed severe mosaic symptoms accompanied by necrosis on the infected leaves. Naju82-S strain is more virulent demonstrated by shorter incubation period and relatively high virus concentration than Naju82-M strain. Five Korean cultivars were tested for their pathogenicity to different strains based on the rate of infection. Results showed that infection rate of cultivars to both strains did not significantly differed from each other. However, under different temperatures, the pathogenicity on the two cultivars such as cultivars Hopumbori and Sessalbori were significantly affected. Hopumbori was moderately resistant to both strains at $10-12^{\circ}C$ and susceptible at $15-18^{\circ}C$. Similarly, Sessalbori was moderately resistant at $10-12^{\circ}C$ to both strains but distinctly differentiated at $15-18^{\circ}C$ wherein it was resistant to mild strain and highly susceptible to severe strain. Other cultivars including Baegdong, Jinyangbori and Neahanssalbori consistently showed susceptible reaction to both strains at varying temperatures tested in this study.

The Attenuation Mechanism and Live Vaccine Potential of a Low-Virulence Edwardsiella ictaluri Strain Obtained by Rifampicin Passaging Culture

  • Shuyi Wang;Jingwen Hao;Jicheng Yang;Qianqian Zhang;Aihua Li
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.167-179
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    • 2023
  • The rifampicin-resistant strain E9-302 of Edwardsiella ictaluri strain 669 (WT) was generated by continuous passage on BHI agar plates containing increasing concentrations of rifampicin. E9-302 was attenuated significantly by 119 times to zebrafish Danio rerio compared to WT in terms of the 50% lethal dose (LD50). Zebrafish vaccinated with E9-302 via intraperitoneal (IP) injection at a dose of 1 × 103 CFU/fish had relative percentage survival (RPS) rates of 85.7% when challenged with wild-type E. ictaluri via IP 14 days post-vaccination (dpv). After 14 days of primary vaccination with E9-302 via immersion (IM) at a dose of 4 × 107 CFU/ml, a booster IM vaccination with E9-302 at a dose of 2 × 107 CFU/ml exhibited 65.2% RPS against challenge with wild-type E. ictaluri via IP 7 days later. These results indicated that the rifampicin-resistant attenuated strain E9-302 had potential as a live vaccine against E. ictaluri infection. A previously unreported amino acid site change at position 142 of the RNA polymerase (RNAP) β subunit encoded by the gene rpoB associated with rifampicin resistance was identified. Analysis of the whole-genome sequencing results revealed multiple missense mutations in the virulence-related genes esrB and sspH2 in E9-302 compared with WT, and a 189 bp mismatch in one gene, whose coding product was highly homologous to glycosyltransferase family 39 protein. This study preliminarily explored the molecular mechanism underlying the virulence attenuation of rifampicin-resistant strain E9-302 and provided a new target for the subsequent study of the pathogenic mechanism of E. ictaluri.