• Title/Summary/Keyword: resistance screening

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Host Range Screening of the Sugar Beet Nematode, Heterodera schachtii Schmidt (사탕무씨스트선충의 기주범위 검정)

  • Kim, Dong Hwan;Cho, Myoung Rae;Yang, Chang Yeol;Kim, Hyeong Hwan;Kang, Taek Jun;Yoon, Jung Beom
    • Korean journal of applied entomology
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    • v.55 no.4
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    • pp.389-403
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    • 2016
  • Sugar beet nematode (Heterodera schachtii Schmidt) was first detected in 2011, in Chinese cabbage grown in the highland areas of Korea. Chemical control of the nematode by nematicides is not feasible due to its cyst-forming characteristics; therefore, the cultivation of non-host crops is a preferable alternative to utilize nematode-infected fields. In this study, a total of 276 plant cultivars belonging to 18 different families were screened to evaluate their resistance to the nematode. Based on the number of cysts formed following nematode inoculation, the tested crops were classified into 3 levels: susceptible, moderately susceptible, and resistant/immune. Among the 276 cultivars tested, 106 cultivars were susceptible, 40 cultivars were moderately susceptible, and 130 cultivars were resistant/immune. Among the resistant/immune cultivars, cyst formation was not observed on eggplant, tomato, lettuce, perilla, carrot, celery, watermelon, oriental melon, cucumber, pumpkin, chives, onion, welsh onion, balloon flower roots, deodeok (Codonopsis lanceolata), Jandae (Adenophora triphylla), and bean. Therefore, these plants are regarded as immune to the cyst nematode. However, many crops belonging to Solanaceae, Asteraceae, Chenopodiaceae, and Poaceae families showed moderate susceptibility or immunity, depending on the crop or cultivar. This study provides a basis for alternative crop recommendations for sugar beet nematode cyst-infected farms in Chinese cabbage production areas.

Improving amber suppression activity of an orthogonal pair of Saccharomyces cerevisiae tyrosyl-tRNA synthetase and a variant of E. coli initiator tRNA, fMam tRNACUA, for the efficient incorporation of unnatural amino acids (효율적인 비천연 아민노산 도입을 위한 효모균 타이로신-tRNA 합성효소와 대장균 시작 tRNA 변이체의 엠버써프레션 활성증가)

  • Tekalign, Eyob;Oh, Ju-Eon;Park, Jungchan
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.420-427
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    • 2018
  • The orthogonal pair of Saccharomyces cerevisiae tyrosyl-tRNA synthetase (Sc YRS) and a variant of E. coli initiator tRNA, fMam $tRNA_{CUA}$ which recognizes the amber stop codon is an effective tool for site-specific incorporation of unnatural amino acids into the protein in E. coli. To evolve the amber suppression activity of the orthogonal pair, we generated a mutant library of Sc YRS by randomizing two amino acids at 320 and 321 which involve recognition of the first base of anticodon in fMam $tRNA_{CUA}$. Two positive clones are selected from the library screening with chloramphenicol resistance mediated by amber suppression. They showed growth resistance against high concentration of chloramphenicol and their $IC_{50}$ values were approximately 1.7~2.3 fold higher than the wild type YRS. In vivo amber suppression assay reveals that mutant YRS-3 (mYRS-3) clone containing amino acid substitutions of P320A and D321A showed 6.5-fold higher activity of amber suppression compared with the wild type. In addition, in vitro aminoacylation kinetics of mYRS-3 also showed approximately 7-fold higher activity than the wild type, and the enhancement was mainly due to the increase of tRNA binding affinity. These results demonstrate that optimization of anticodon recognition by engineered aminoacyl tRNA synthetase improves the efficiency of unnatural amino acid incorporation in response to nonsense codon.

Development of Screening Method for the Soluble Recombinant Protein using β-Lactamase as a Fusion Partner (β-Lactamase 접합 단백질 발현 시스템을 이용한 가용성 재조합 단백질 탐색 기술 개발)

  • Lee, Jae-Hun;Hwang, Bum-Yeol;Kim, Byung-Gee;Lee, Sun-Gu
    • Korean Chemical Engineering Research
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    • v.47 no.5
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    • pp.624-629
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    • 2009
  • It is the most important step to screen soluble and insoluble proteins when we attempt to improve the solubility of recombinant proteins through directed evolution approach. Here we show that the solubility of a recombinant protein in vivo can be examined by expressing the recombinant protein with beta-lactamase as a fusion partner. First we constructed an expression system which can produc a fusion protein with the C-terminal of beta-lactamase. Two soluble proteins, i.e. adenine deaminase and aspartate aminotransferase, and insoluble GlcNAc-2-epimerase were cloned into the developed expression vector, respectively. We investigated the effect of the expression of the three recombinant fusion proteins on the growth of E. coli, and confirmed that the solubilities of the recombinant proteins correlated with cell growth rates.

A Case of Vancomycin-Resistant Enterococcal Sepsis in Neonate (신생아에서 Vancomycin 내성 장구균 감염 1례)

  • Bae, Soo Jung;Choi, Gui Jean;Kim, Chun Soo;Lee, Sang Lak;Kim, Heung Sik;Kang, Chin Moo
    • Pediatric Infection and Vaccine
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    • v.6 no.2
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    • pp.261-266
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    • 1999
  • Vancomycin-resistant enterococcus(VRE) was first isolated from various specimens of patients with renal failure or leukemia in 1988. Thereafter VRE has been increasing gradually and became one of the clinically important palhogenic organisms currently. We experienced a case of E. faecalis sepsis in a 4 day old neonate. She was born at 39 weeks gestational age with 2,900gm weight by Cesarean section delivery due to breech presentation. She had had swelling and motion limitation of the left knee joint with fever for one day at age of 4 day and was transfered to our hospital. Ultrasonographic examination of her left knee joint showed some inflammatory change. E. faecalis was isolated from the blood. The organism showed resistance to vancomycin on drug susceptibility test using BHI agar screening test and disk diffusion method. After treatment with ampicillin-sulbactam for 3 weeks the baby was improved. Although VRE infection has been considered rare in Korea. considerable number of demonstrative studies about VRE isolation have been reported recently thus adequate countermeasures are needed to reduce the emergence and prevent nosocomial spreading of this organism.

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Effects of Soil Textures on Infectivity of Root-Knot Nematodes on Carrot

  • Kim, Eunji;Seo, Yunhee;Kim, Yong Su;Park, Yong;Kim, Young Ho
    • The Plant Pathology Journal
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    • v.33 no.1
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    • pp.66-74
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    • 2017
  • This study was conducted to examine infectivity (penetration and gall and egg-mass formations) of the root-knot nematodes, Meloidogyne incognita and M. hapla, on carrots grown in soil conditions of 5 different soil textures consisting of bed-soil (b) and sand (s) mixtures (b-s mixtures) at the ratios of 10:0, 7:3, 5:5, 3:7, and 0:10. For M. incognita, the nematode penetration rates in b-s of 0:10 (100% sand) were significantly higher than in the other b-s mixtures, more greatly at 2 and 5 days after inoculation than at 10 DAI, while no significant differences in the penetration rates were mostly shown for M. hapla at the above DAI. However, for both nematodes, gall and egg-mass formations were remarkably increased in the b-s mixture of 0:10, compared to the other b-s mixtures, which is coincided with the general aspects of severe nematode infestations in sandy soils. This suggests the increased gall and egg-mass formations of M. incognita should be derived from the increased penetration rates in the sandy soil conditions, which provide a sufficient aeration due to coarse soil nature for the nematodes, leading to their mobility increased for the enhanced root penetration. For M. hapla, it is suggested that the sandy soil conditions affect positively on the healthy plant growth with little accumulation of the inhibitory materials and sufficient aeration, enhancing the nematode growth and feeding activities. All of these aspects provide information reliable for the development screening techniques efficient for the evaluation of the nematode resistance in the breeding programs.

Screening of Plant Extracts and Identification of their Insecticidal Metabolites against Myzus persicae (복숭아혹진딧물 방제용 식물추출물 탐색 및 살충성분 구명)

  • Yang, Si young;Lim, Da jung;Kim, Yeo Hee;Kim, In Seon
    • Korean Journal of Environmental Agriculture
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    • v.37 no.2
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    • pp.125-134
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    • 2018
  • BACKGROUND: Green peach aphid (Myzus persicae) is an insect pest that significantly affects crop production. A number of pesticides have been used for aphid control, but their concerns on insect resistance and food safety have required alternative methods for pest management. In an effort to find for an alternative approach to aphid control, we screened plants extracts and examined their potentiality as insecticidal bio-resources. METHODS AND RESULTS: Two hundred and ninety eight plant extracts were examined for insecticidal activity against the aphid, and the best candidate among them was chosen for further study. The extracts from Cinnamomum camphora was determined to be the best candidate exhibiting insecticidal activity more than 60% at a level of $1,000{\mu}g/mL$. GC/MS analyses detected camphor, borneol, 4-terpineol, ${\alpha}$-terpineol and caryophyllene oxide as major compositions from the extracts obtained by hydrodistillation. Caryophyllene oxide exhibited the highest insecticidal activity with a $LC_{50}$ value of $237{\mu}g/mL$. Camphor lowered significantly the $LC_{50}$ value of caryophyllene oxide and increased largely its concentration in aphid, suggesting that camphor played a role in enhancing the insecticidal activity of caryophyllene oxide. CONCLUSION: This study suggested that camphor and caryophyllene oxide may be used as an insecticidal bio-resource for insect control against green peach aphid.

Toll-like Receptor 4 Polymorphism and Periodontitis in Korean Population

  • Park, Ok-Jin;Shin, Seung-Yun;Chung, Chong-Pyoung;Ku, Young;Choi, Young-Nim;Kim, Kack-Kyun
    • International Journal of Oral Biology
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    • v.31 no.1
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    • pp.1-6
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    • 2006
  • The primary cause of periodontitis is plaque-associated anaerobic gram-negative bacteria. As shown in the patients with defects in the number or function of neutrophils, innate immunity plays an important role in resistance to bacterial infection and periodontitis. Toll-like receptor 4(TLR4) is one of the key receptors that recognize the molecular patterns of microbes and initiate innate immune response. To understand the role of TLR4 in the pathogenesis of periodontitis, we investigated whether Asp299Gly of TLR4 mutation is associated with periodontitis in Korean population. Subjects for this study included 90 healthy subjects and 98 periodontitis patients. The Asp299Gly mutation was screened by PCR-Restriction Fragment Length Polymorphism(RFLP) of genomic DNA from blood cells using a primer that creates a NcoI restriction site only in the mutant allele. The Asp299Gly mutation was not found in all subjects tested. Our results suggest that the Asp299Gly mutation of TLR4 is very rare in a Korean population. Further mutation screening may be required to determine the role of TLR4 in the pathogenesis of periodontitis.

A Study on the Thermal Design for A Signal Processor in the Micro-Wave Seeker (초고주파 탐색기 신호처리부의 방열설계에 관한 연구)

  • Lee, Won-Hee;Yu, Young-Joon;Kim, Ho-Yong
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.39 no.1
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    • pp.76-83
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    • 2011
  • This paper focuses on the thermal design of a signal processor in Micro-Wave Seeker. High temperature environment and ESS(Environmental Stress Screening) test condition should be considered in designing a signal processor. First, we performed the thermal analysis to know conditions under which a signal processor is thermally reliable. As a result of thermal analysis, we found that adopting heat transfer block to the thermally fragile components is most efficient, because the heat transfer block can control the thermal loads of the individual components. Next, we verified this solution by numerical simulation and experiment and concluded that thermal reliability of a signal processor can be achieved. Maximum temperature difference between numerical simulation and experiment is about $2^{\circ}C$.

Isolation and Characterization of the Colletotrichum acutatum ABC Transporter CaABC1

  • Kim, Suyoung;Park, Sook-Young;Kim, Hyejeong;Kim, Dongyoung;Lee, Seon-Woo;Kim, Heung Tae;Lee, Jong-Hwan;Choi, Woobong
    • The Plant Pathology Journal
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    • v.30 no.4
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    • pp.375-383
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    • 2014
  • Fungi tolerate exposure to various abiotic stresses, including cytotoxic compounds and fungicides, via their ATP-driven efflux pumps belonging to ATP-binding cassette (ABC) transporters. To clarify the molecular basis of interaction between the fungus and various abiotic stresses including fungicides, we constructed a cDNA library from germinated conidia of Colletotrichum acutatum, a major anthracnose pathogen of pepper (Capsicum annum L.). Over 1,000 cDNA clones were sequenced, of which single clone exhibited significant nucleotide sequence homology to ABC transporter genes. We isolated three fosmid clones containing the C. acutatum ABC1 (CaABC1) gene in full-length from genomic DNA library screening. The CaABC1 gene consists of 4,059 bp transcript, predicting a 1,353-aa protein. The gene contains the typical ABC signature and Walker A and B motifs. The 5'-flanking region contains a CAAT motif, a TATA box, and a Kozak region. Phylogenetic and structural analysis suggested that the CaABC1 is a typical ABC transporter gene highly conserved in various fungal species, as well as in Chromista, Metazoans, and Viridiplantae. We also found that CaABC1 was up-regulated during conidiation and a minimal medium condition. Moreover, CaABC1 was induced in iprobenfos, kresoxim-methyl, thiophanate-methyl, and hygromycin B. These results demonstrate that CaABC1 is necessary for conidiation, abiotic stress, and various fungicide resistances. These results will provide the basis for further study on the function of ABC transporter genes in C. acutatum.

Isolation and Sequencing of the cDNA Encoding ${\beta}-tubulin$ from Pleurotus sajor-caju (여름느타리버섯으로부터 ${\beta}-tubulin$ cDNA의 분리 및 염기서열 결정)

  • Kim, Beom-Gi;Shin, Pyung-Gyun;Jeong, Mi-Jeong;Park, Soo-Chul;Yoo, Young-Bok;Ryu, Jin-Chang;Kwon, Suk-Tae
    • The Korean Journal of Mycology
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    • v.25 no.1 s.80
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    • pp.1-5
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    • 1997
  • The cDNA encoding ${\beta}-tubulin$ of Pleurotus sajor-caju was isolated using an internal gene segment probe amplified by polymerase chain reaction (PCR) of genomic DNA and by cDNA library screening. The cDNA was consisted of 1560 nucleotides(nt), including a 5'-untranslation region (UTR) of 27nt, an open reading frame (ORF) of 1341nt, and a 3'-UTR of 191nt. The ORF encoded a protein of 446 amino acids(aa), which shows over 80% homology with ${\beta}-tubulins$ of other filamentous fungi. Southern hybridization analysis showed that there were two isotypes of ${\beta}-tubulin$ genes in P. sajor-caju. Through sequence analysis we found that ${\beta}-tubulin$ had a unusual $Cys^{165}$ residue, which might be a significant factor for the insensitivity of fungi to fungicide benomyl.

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