• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.69.1-69
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• 2003

RSH (relA/spoT homolog) has been known to determine the level of guanosine tetraphosphate (ppGpp) and guanosine pentaphosphate (pppGpp), which are the effector nucleotide of the prokaryotic stringent response and also play a role in antibiotic production and differentiation in Streptomyces species but not a little in eukaryotic organism, especially in plant. Salicylic acid (SA), a critical signal molecule of establishing systemic acquired resistance (SAR), could induce SAR in Pepper (Capcicum annuum) against Phytophthora capsici. And the extent of SAR induction was in proportion to the dosage of SA (or BTH). Suppression subtractive hybridization (SSH), a PCR-based method for cDNA subtraction, was carried out between SA-treated and non-SA-treated pepper leaves to isolate genes which may be responsible for defense signaling against pathogens. Early upregulated gene was selected from reverse northern and kinetics of SSH-genes transcripts in SA-treated pepper leaves upon SA treatment. Full-length cDNA of the gene (PepRSH； Pepper RelA / SpoT homolog) had an open reading frame (ORF) of 2166 bp encoding a protein of 722 amino acids and a significant homology with (p)ppGpp phosphohydrolase or synthetase. Genomic DNA gel blot analysis showed that pepper genome has at least single copy of PepRSH. PepRSH transcripts was very low in untreated pepper leaves but strongly induced by SA and methyljasmonic acid (MeJA), indicating that PepRSH may share common SA and MeJA-mediated signal transduction pathway Functional analysis in E. coli showed PepRSH confers phenotypes associated with (p)ppGpp synthesis through a complementation using active site mutagenesis.

• Fisheries and Aquatic Sciences
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• 제23권4호
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• pp.7.1-7.8
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• 2020

Background: To minimize the use of antibiotics and to obtain a more sustainable fish culture and aquaculture industry, development of alternative natural source of immunostimulant to replace antibiotic in aquafeed is highly needed. Objective: Dietary inclusion effect of yacon (YC), ginger (GG), and blueberry (BB) on growth, body composition, and disease resistance of black rockfish against Vibrio anguillarum was compared to ethoxyquin (EQ). Methods: Four hundred eighty juvenile (an initial weight of 4.2 g) fish were randomly distributed into 12 of 50 L flowthrough tanks (forty fish per tank). Four experimental diets were prepared; the control (Con) diet with 0.01% EQ inclusion, and YC, GG, and BB diets at 1% each additive inclusion. Each additive was included into the experimental diets at the expense of wheat flour. Each diet was assigned to triplicate tanks of fish and hand-fed to satiation twice daily for 8 weeks. At the end of 8-week feeding trial, 20 fish from each tank fish were artificially infected by intraperitoneal injection with 0.1 mL of culture suspension of pathogenic V. anguillarum containing 3.3 × 10⁶ cfu/mL respectively. Fish were monitored for the following 8 days after V. anguillarum infection and dead fish were removed every 6 h for the first 4 days and 12 h for the rest of the study. Results: Weight gain, specific growth rate (SGR), and feed efficiency ratio (FER) of fish fed the YC diet was higher than those of fish fed all other diets. However, feed consumption, protein efficiency ratio, and protein retention was not affected by dietary additive. Moisture, crude protein, and crude lipid content of the whole body of fish were affected by dietary additive. Analysis of the Kaplan-Meier survival curves showed that survival of fish fed the YC, BB, and GG diets was higher than the Con diet. Conclusion: Oral administration of YC can improve not only weight gain, SGR, and FER of black rockfish, but lower mortality of rockfish at occurrence of V. anguillarum.

• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.1057-1062
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• 2004

The entomopathogenic bacterium Bacillus thuringiensis is the most widely used biopesticide. Insecticidal proteins, coded by genes located in plasmids, form typical parasporal, crystalline inclusions during sporulation. We isolated a Bacillus thuringiensis strain having insecticidal activity against larvae of the house fly (M. domestica) from the soils at a pig farm in Korea, and named it Bacillus thuringiensis SM. The culture filtrate from Bacillus thuringiensis SM showed strong lethality (83.3%) against M. domestica larvae. The parasporal crystal is enclosed within the spores' outermost envelope, as determined by transmission electron microscopy, and exhibited a bipyramidal form. The crystal proteins of strain SM consisted of five proteins with molecular weights of approximately ~130, ~80, ~68, ~42, and ~27 kDa on a 10% SDS-PAGE (major band, a size characteristic of Cry protein). Examination of antibiotic resistance revealed that the strain SM showed multiple resistant. The strain SM had at least three different plasmids with sizes of 6.6, 9.3, and 54 kb. Polymerase chain reactions (PCRs) revealed the presence of cry1, cry4A2, and cry11A1 genes in the strain SM. The cry1 gene profile of the strain SM appeared in the three respective products of 487 bp [cry1A(c)], 414 bp [cry1D], and 238 bp [cry1A(b)]. However, the strain SM has not shown the cry4A2 md cry11A1 genes. In in vivo toxicity assays, the strain SM showed high toxicity on fly larvae (M. domestic) [with $LC_{50}$ of 4.2 mg/ml, $LC_{90}$ of 8.2 mg/ml].

• Clinical and Experimental Pediatrics
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• 제54권4호
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• pp.146-151
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• 2011

Streptococcus pneumoniae remains a leading cause of invasive infections including bacteremia and meningitis, as well as mucosal infections such as otitis media and pneumonia among children and adults. The 7-valent pneumococcal conjugate vaccine (PCV7) was licensed for use among infants and young children in many countries including Korea. The routine use of PCV7 has resulted in a decreased incidence of invasive pneumococcal disease (IPD) by the vaccine serotypes among the vaccinees and substantial declines in IPD among unvaccinated populations such as older children and adults as well. In addition, there are increasing evidences to suggest that routine immunization with PCV7 is changing the epidemiology of pneumococcal diseases such as serotype distribution of IPD, nasopharyngeal colonization, and antibiotic resistance patterns. In contrast, there is an increase in the number of IPDs caused by nonvaccine serotypes, though it is much smaller than overall declines of vaccine serotype diseases. Several vaccines containing additional serotypes have been developed and tested clinically in order to expand the range of serotypes of Streptococcus pneumoniae. Recently two new pneumococcal protein conjugate vaccines, 10-valent pneumococcal conjugate vaccine (PCV10) and 13-valent pneumococcal conjugate vaccine (PCV13), have been approved for use in several countries including Korea. This report summarizes the recommendations approved by the Committee on Infectious Diseases, the Korean Pediatric Society.

• Bulletin of the Korean Chemical Society
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• 제32권8호
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• pp.2695-2699
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• 2011

${\beta}$ Ketoacyl acyl carrier protein synthase III (KAS III) initiates fatty acid synthesis in bacteria and is a key target enzyme to overcome the antibiotic resistance problem. In our previous study, we found flavonoid inhibitors of Enterococcus faecalis KAS III and proposed three potent antimicrobial flavonoids against Enterococcus faecalis and Vancomycin-resistant Enterococcus faecalis with MIC values in the range of 128-512 ${\mu}g/mL$ as well as high binding affinities on the order from $10^6$ to $10^7\;M^{-1}$. Using these series of flavonoids, we conducted biological assays as well as docking study to find potent flavonoids inhibitors of Staphylococcus aureus KAS III with specificities against Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. Here, we propose that naringenin (5,7,4'-trihydroxyflavanone) and eriodictyol (5,7,3',4'-tetrahydroxyflavanone) are potent antimicrobial inhibitors of Staphylococcus aureus KAS III with binding affinity of $3.35{\times}10^5$ and $2.01{\times}10^5\;M^{-1}$, respectively. Since Arg38 in efKAS III is replaced with Met36 in saKAS III, this key difference caused one hydrogen bond missing in saKAS III compared with efKAS III, resulting in slight discrepancy in their binding interactions as well as decrease in binding affinities. 4'-OH and 7-OH of these flavonoids participated in hydrogen bonding interactions with backbone carbonyl of Phe298 and Ser152, respectively. In particular, these flavonoids display potent antimicrobial activities against various MRSA strains in the range of 64 to 128 ${\mu}M$ with good binding affinities.

• Journal of Microbiology and Biotechnology
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• 제33권2호
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• pp.211-218
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• 2023

A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Em^r), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.

• 한국임상수의학회지
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• 제32권2호
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• pp.191-195
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• 2015

7년령, 중성화 수컷 고양이가 무기력과 식욕부진으로 내원하였다. 실험실 검사는 말초혈액 호중구에서 중등도의 퇴행성 변화, free 및 total thyroxine의 농도 감소 및 혈액배양에서 세균증식이 관찰되었다. 16S ribosomal RNA 및 heat shock protein 60 유전자 검사 결과 세균은 Enterobacter cloacae로 확인되었다. 항생제 최소억제농도 평가결과 16개 항생제에 대한 다약제 내성이 관찰되었다. 중합효소연쇄반응 및 시퀀싱 결과 $bla_{TEM-1}$, $bla_{CTX-M-15}$ 및 플라스미드 유래 $bla_{ACT-1}$ 유전자에 대한 양성 반응이 관찰되었으며, 결과적으로 확인된 세균이 광범위 beta-lactamase 및 ampC beta-lactamase를 합성하는 플라스미드를 보유한 것을 확인하였다. 환자는 1개월 간 항생제와 levothyroxine 치료를 받은 후 증상이 호전되었고, 치료 후 갑상선 기능검사와 혈액배양 결과 이상소견은 소실되었다. 이 증례는 고양이의 지역사회획득 다약제내성 E. cloacae에 의한 정상 갑상샘 질환 증후군의 첫 예이다. 신속한 진단과정과 적절한 항생제 선택에 의해 이환된 고양이는 패혈증으로부터 회복되었다.

• Journal of Microbiology and Biotechnology
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• 제30권4호
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• pp.526-532
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• 2020

A bacterial strain, designated B301^T and isolated from raw chicken meat obtained from a local market in Korea, was characterized and identified using a polyphasic taxonomic approach. Cells were gram-negative, non-motile, obligate-aerobic coccobacilli that were catalase-positive and oxidase-negative. The optimum growth conditions were 30℃, pH 7.0, and 0% NaCl in tryptic soy broth. Colonies were round, convex, smooth, and cream-colored on tryptic soy agar. Strain B301^T has a genome size of 3,102,684 bp, with 2,840 protein-coding genes and 102 RNA genes. The 16S rRNA gene analysis revealed that strain B301^T belongs to the genus Acinetobacter and shares highest sequence similarity (97.12%) with A. celticus ANC 4603^T and A. sichuanensis WCHAc060041^T. The average nucleotide identity and digital DNA-DNA hybridization values for closely related species were below the cutoff values for species delineation (95-96% and 70%, respectively). The DNA G+C content of strain B301^T was 37.0%. The major respiratory quinone was Q-9, and the cellular fatty acids were primarily summed feature 3 (C_16:1 ω6c/C_16:1 ω7c), C_16:0, and C_18:1 ω9c. The major polar lipids were phosphatidylethanolamine, diphosphatidyl-glycerol, phosphatidylglycerol, and phosphatidyl-serine. The antimicrobial resistance profile of strain B301^T revealed the absence of antibiotic-resistance genes. Susceptibility to a wide range of antimicrobials, including imipenem, minocycline, ampicillin, and tetracycline, was also observed. The results of the phenotypic, chemotaxonomic, and phylogenetic analyses indicate that strain B301^T represents a novel species of the genus Acinetobacter, for which the name Acinetobacter pullorum sp. nov. is proposed. The type strain is B301^T (=KACC 21653^T = JCM 33942^T).

• Applied Biological Chemistry
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• 제37권4호
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• pp.221-225
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• 1994

농산폐섬유소 자원을 이용하여 세균 단세포단백질(single cell protein)을 생산하기 위한 연구로서 부엽토에서 분리된 Cellulomonas sp. KL-6의 증식에 미치는 열처리 및 항생물질의 효과에 대해 조사하였다. KL-6 균주는 $65^{\circ}C$에서 5분, $75^{\circ}C$에서 1분 및 $85^{\circ}C$에서 15초까지 열처리시에는 증식하였고 그 이상의 온도에서는 처리시간에 관계없이 자라지 못하였다. $48^{\circ}C$에서 10분간의 예비 열처리는 본 균주의 열저항성을 증가시키지 못하였다. 기질 중 casamino acid(1%), yeast extract(1%), xylose(5%)는 KL-6 균주의 열저항성을 증가시켰으나 cysteine(0.03 M), sodium citrate(1%), casein(1%)은 반대로 열저항성을 감소시켰다. KL-6 균주는 $penicillin-G(1.536\;{\mu}g/ml)$와 $ampicillin(3.125\;{\mu}g/ml)$에 감수성인 반면, kanamycin에 대해서는 $200\;{\mu}g/ml$ 이상에서도 강한 내성을 보였다. SDS 처리에 의해 kanamycin의 내성이 제거되었고 제거율은 $9.2{\sim}31.2%$로 각각 나타났다.

• 생명과학회지
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• 제24권7호
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• pp.769-776
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• 2014

Anisomycin은 Streptomyces griseolus에 의하여 생성되는 항생제의 일종으로 flagecidin으로도 알려져 있으며, ribosomal 28S subunit에 결합함으로서 단백질의 생성을 억제하는 것으로 알려져 있다. TRAIL은 ligand로서의 death receptor와의 결합을 통하여 세포의 apoptosis를 유발하는 것으로 알려져 있으나, 많은 암세포에서는 이미 TRAIL에 대한 저항성을 획득하여 TRAIL 유도 apoptosis를 회피하는 능력을 가지고 있다. 본 연구에서는 TRAIL 저항성 Hep3B 간암세포를 대상으로 anisomycin이 TRAIL 매개 apoptosis를 촉진 시킬 수 있는지의 여부를 조사하였다. 본 연구의 결과에 의하면, 단독 처리 조건에서 Hep3B 세포의 증식에 유의적인 영향을 미치는 않았던 anisomycin과 TRAIL의 동시 처리는 anisomycin 처리 농도 의존적으로 세포의 증식을 시켰으며, 이는 caspase 활성화를 통한 apoptosis 유발 증가와 연관성이 있음을 확인하였다. 특히 siRNA를 이용한 Hep3B 세포의 인위적인 Bid 발현의 차단은 anisomycin과 TRAIL 동시 처리군에 비하여 apoptosis 유발능이 더욱 증대시켜 TRAIL 연관 Bid의 truncation을 통한 미토콘드리아 의존적 apoptosis 유발 과정을 anisomycin이 효과적으로 촉진시켰음을 보여주었다. 따라서 본 연구의 결과는 anisomycin과 TRAIL의 동시 처리는 TRAIL 저항성 암세포의 사멸을 촉진시킬 수 있는 효과적인 방법임을 의미한다.