• Title/Summary/Keyword: DNA homology

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First Report on Isolation of Salmonella Enteritidis from Eggs at Grocery Stores in Korea

  • Kim, Young Jo;Song, Bo Ra;Lim, Jong Su;Heo, Eun Jeong;Park, Hyun Jung;Wee, Sung Hwan;Oh, Soon Min;Moon, Jin San
    • Food Science of Animal Resources
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    • v.33 no.2
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    • pp.239-243
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    • 2013
  • Salmonella Enteritidis is responsible for causing foodborne diseases upon consumption of egg products. While cases of S. Enteritidis isolation from eggs have been reported in other countries, no such cases have previously been reported in Korea. In this study, we report the first isolation and identification of S. Enteritidis from domestically distributed eggs in Korea. Eggs were collected from eight countrywide grocery stores during different seasons between 2011 and 2012. Egg contents and washing solution of egg shells were incubated in buffered peptone water, and the enriched broth was further enriched in tetrathionate broth and Rappaport-Vassiliadis. The secondary enriched broth was streaked on xylose lysine desoxycholate agar. The suspected colonies were confirmed to S. Enteritidis by a biochemical test, serotyping, and PCR test. Genetic relatedness among the isolates was analyzed using Diversilab Salmonella kit. Three strains of S. Enteritidis were isolated from egg contents and egg shells collected from grocery stores of the Eumseong-city in the fall of 2011. All three stains showed resistance to chloramphenicol, streptomycin, nalidixic acid, and ampicillin by the disk diffusion method. In addition, the isolates showed more than 99% DNA homology, indicating that they were presumably identical strains. Therefore, there is a requirement to monitor and control against S. Enteritidis from eggs in Korea.

Expression of Neurotensin/Neuromedin N Precursor in Murine Mast Cells

  • Ahn, Hyun-Jong;Cho, Jeong-Je
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.6
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    • pp.495-501
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    • 2001
  • We have cloned the mouse neurotensin/neuromedin N (NT/N) gene from the murine mast cell line Cl.MC/C57.1 for the first time. The murine NT/N cDNA clone consisted of 765 nucleotides and coded for 169 peptide residues with an N-terminal signal peptide, and the C-terminal region contained of one copy of neurotensin (NT) and one copy of neuromedin N (NN). Total of four Lys-Arg dibasic motifs were present; one each at the middle of the open reading frame, at the N-terminal of NN, at the C-terminal of NT, and between NN and NT. Amino acid sequence analysis of the mouse NT/N revealed 90% homology to that of the rat NT/N gene. NT/N is expressed in murine mast cell lines (Cl.MC/C57.1 and P815), but not in murine bone marrow-derived mast cells (BMMCs), murine macrophage cell line (RAW 264.7), nor in murine T cell line (EL-4). NT/N mRNA in C1.MC/C57.1 is highly inducible by IgE cross-linking, phorbol myristate acetate, neurotensin, and substance P. Following the treatment of demethylating agent, 5-azacytidine (5-azaC), the NT/N gene was induced in BMMCs in response to IgE cross-linking. 5-azaC-treated BMMCs did not express the NT/N gene without additional stimuli. These findings suggested that the regulation of NT/N gene expression was dependent on the effects of not only gene methylation but also enhancer and/or repressor proteins acting on the NT/N promoter.

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Triclosan Resistance in a Bacterial Fish Pathogen, Aeromonas salmonicida subsp. salmonicida, is Mediated by an Enoyl Reductase, FabV

  • Khan, Raees;Lee, Myung Hwan;Joo, Haejin;Jung, Yong-Hoon;Ahmad, Shabir;Choi, Jinhee;Lee, Seon-Woo
    • Journal of Microbiology and Biotechnology
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    • v.25 no.4
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    • pp.511-520
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    • 2015
  • Triclosan, the widely used biocide, specifically targets enoyl-acyl carrier protein reductase (ENR) in the bacterial fatty acid synthesis system. Although the fish pathogen Aeromonas salmonicida subsp. salmonicida exhibits triclosan resistance, the nature of this resistance has not been elucidated. Here, we aimed to characterize the triclosan resistance of A. salmonicida subsp. salmonicida causing furunculosis. The fosmid library of triclosan-resistant A. salmonicida subsp. salmonicida was constructed to select a fosmid clone showing triclosan resistance. With the fosmid clone showing triclosan resistance, a subsequent secondary library search resulted in the selection of subclone pTSR-1. DNA sequence analysis of pTSR-1 revealed the presence of a chromosomal-borne fabV-encoding ENR homolog. The ENR of A. salmonicida (FabVas) exhibited significant homology with previously known FabV, including the catalytic domain YX(8)K. fabVas introduction into E. coli dramatically increased its resistance to triclosan. Heterologous expression of FabVas might functionally replace the triclosan-sensitive FabI in vivo to confer E. coli with triclosan resistance. A genome-wide search for fabVas homologs revealed the presence of an additional fabV gene (fabVas2) paralog in A. salmonicida strains and the fabVas orthologs from other gram-negative fish pathogens. Both of the potential FabV ENRs expressed similarly with or without triclosan supplement. This is the first report about the presence of two potential FabV ENRs in a single pathogenic bacterium. Our result suggests that triclosan-resistant ENRs are widely distributed in various bacteria in nature, and the wide use of this biocide can spread these triclosan-tolerant ENRs among fish pathogens and other pathogenic bacteria.

Gibberellin-Producing Endophytic Fungi Isolated from Monochoria vaginalis

  • Ahmad, Nadeem;Hamayun, Muhammad;Khan, Sumera Afzal;Khan, Abdul Latif;Lee, In-Jung;Shin, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1744-1749
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    • 2010
  • The role of endophytic fungi in plant growth and development is well documented. However, endophytic fungi with growth promotion capacity have never been isolated from weeds previously. In the current study, we isolated 8 fungal endophytes from the roots of Monochoria vaginalis, a serious weed of rice paddy in Korea. These isolates were screened on Waito-C, in order to identify plant growth promoting metabolites. Two fungal isolates (M5.A & M1.5) significantly promoted the plant height and shoot length of Waito-C during preliminary screening experiments. The culture filtrates (CFs) of M5.A and M1.5 also promoted the shoot length of Echinocloa crusgalli. Gibberellins (GAs) analysis of the CFs of M5.A and M1.5 showed that these endophytic fungi secrete higher quantities of GAs as compared with wild-type G. fujikuroi KCCM12329. The CF of M5.A contained bioactive GAs ($GA_3$, 2.8 ng/ml; $GA_4$, 2.6 ng/ml, and $GA_7$, 6.68 ng/ml) in conjunction with physiologically inactive $GA_9$ (1.61 ng/ml) and $GA_{24}$ (0.18 ng/ml). The CF of M1.5 contained physiologically active GAs ($GA_3$, 1.64 ng/ml; $GA_4$, 1.37 ng/ml and $GA_7$, 6.29 ng/ml) in conjunction with physiologically inactive $GA_9$ (3.44 ng/ml), $GA_{12}$ (0.3 ng/ml), and $GA_{24}$ (0.59 ng/ml). M5.A and M1.5 were identified as new strains of Penicillium sp. and Aspergillus sp., respectively, based on their 18S rDNA sequence homology and phylogenetic analysis.

Comparative Genomics Study of Interferon-$\alpha$ Receptor-1 in Humans and Chimpanzees

  • Kim, Il-Chul;Chi, Seung-Wook;Kim, Dae-Won;Choi, Sang-Haeng;Chae, Sung-Hwa;Park, Hong-Seog
    • Genomics & Informatics
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    • v.3 no.4
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    • pp.142-148
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    • 2005
  • The immune response-related genes have been suggested to be the most favorable genes for positive selection during evolution. Comparing the entire DNA sequence of chimpanzee chromosome 22 (PTR22) with human chromosome 21 (HSA21), we have identified 15 orthologs having indel in their coding sequences. Among them, interferon-${\alpha}$ receptor-1 gene (IFNAR1), an immuneresponse-related gene, is subjected to comparative genomic analysis. Chimpanzee IFNAR1 showed the same genomic structure as human IFNAR1 (11 exons and 10 introns) except the 3 bp insertion in exon 4. The sequence alignment of IFNAR1 coding sequence indicated that 'ISPP' amino acid sequence motif is highly conserved in chimpanzee and other animals including mouse and chicken. However, the human IFNAR1 shows that one proline residue is missing in the sequence motif. The homology modeling of the IFNAR1 structures suggests that the proline deletion in human IFNAR1 leads to the formation of the following ${\alpha}$-helix, whereas two sequential prolines in chimpanzee IFNAR1 inhibit it. As a result, human IFNAR1 may adopt a characteristic structure distinct from chimpanzee IFNAR1. This human specific trait could contribute to specific immune response in the most optimized manner for humans. Further molecular biological studies on the IFNAR1 will help us to gain insights into the molecular implication of species-specific host-pathogen interaction in primate evolution.

Sturcture of the Rice Glutelin Multigene Family and Its Expression (쌀 Glutelin 유전자군의 구조 및 발현조절)

  • 황영수
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.261-282
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    • 1987
  • Plants store a significant amount of their nitrogen, sulfur and carbon reserves as storage proteins in seed tissues. The major proteins present in rice seeds are the glutelins. Glutelins are initially synthesized at 4-6 days postanthesis and deposited into protein bodies via Golgi apparatus. Based on nucleic acid sequences and Southern blot analysis, the three isolated glutelin genomic clones were representative members of three gene subfamilies each containing 5 to 8 copies. A comparison of DNA sequences displayed by relevant regions of these genomic clones showed that two subfamilies, represented by clones, Gt1 and Gt2, were closely, related and probably evolved by more recent gene duplication events. The 5' flanking and coding sequences of Gt1 and Gt2 displayed at least 87% homolgy. In contrast, Gt3 showed little or no homolgy in the 5' flanking sequences upstream of the putative CAAT boxes and exhibited significant divergence in all other portions of the gene. Conserved sequences in the 5' flanking regions of these genes were identified and discussed in light of their potential regulatory role. The derived primary sequences of all three glutelin genomic clones showed significant homology to the legume 11S storage proteins indicating a common gene origin. A comparison of the derived glutelin primary sequences showed that mutations were clustered in three peptide regions. One peptide region corresponded to the highly rautable hypervariable region of legume peptide region of legume 11S storage proteins, a potential target area for protein modification. Expression studies indicated that glutelin mRNA transcripts are differentially accumulated during endosperm development. Promoterss of Gt2 and Gt3 were functional as they direct transient expression of chloramphenicol acetyltransferase in cultured plant cell.

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Identification and expression of leuD Gene in Rice (Oryza sativa L.) (벼(Oryza sativa L.)의 leuD 유전자)

  • Lee, Eun-Tag;Kang, Sang-Gu
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.772-777
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    • 2007
  • A rice OsLeuD gene for small subunit of 3-isopropylmalate isomerase (IPMI) (EC 4.2.1.33) has been isolated. OsLeuD gene is located on 109.3 cM of chromosome 2. OsLeuD gene was expressed abundantly in metabolically active organs including leaves and developing seeds, indicating that OsLeuD gene expression is developmentally regulated. The cDNA of OsLeuD gene was coded for 257 amino acids which showed 58% and 48% homology to small subunits of IPMI in OsLeuD genes of cyanobacteria and green sulfur bacteria, respectively. The molecular character of OsLeuD is closely related to those of photosynthetic bacteria rather than those of eukaryotes including fungi and yeast. This suggests that OsLeuD gene in chromosomal genome of plants may possibly be originated from chloroplast genome.

Expression of the Galactokinase Gene (gaIK) from Lactococcus lactis asp. lactis ATCC7962 in Escherichia coil

  • Lee, Hyong-Joo;Lee, Jung-Min;Park, Jae-Yeon;Lee, Jong-Hoon;Kim, Jeong-Hwon;Chang, Hea-Choon;Chung, Dae-Kyun;Kim, Somi-Cho
    • Journal of Microbiology
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    • v.40 no.2
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    • pp.156-160
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    • 2002
  • The whole gal/lae operon genes of Lactococcus lactis ssp. lactis 7962 were reported as follows: galA-galM-galK-galT-lacA -lacZ-galE. The galK gene encoding a galactokinase involved in one of the Leloir pathways for galactose metabolism was found to be 1,197 bp in length and encodes a protein of 43,822 Da calculated molecular mass. The deduced amino acid sequence showed over 50% homology with GaIK proteins from several other lactic acid bacteria. The galK gene was expressed in E. coli and the product was identified as a 43 kDa protein which corresponds to the estimated size from the DNA sequence. The galactokinase activity of recombinant 5. coli was about 8 times greater against that of the host strain and more than 3 times higher than the induced L. lactis 7962.

The polymorphism of Theileria buffeli major surface protein associate with their clinical signs in holstein in Korea (국내 젖소에서 Theileria buffeli 주요 표면 단백질 유전자의 다양성 분석)

  • Yu, Do-Hyeon;Li, Ying-Hua;Chae, Joon-Seok;Park, Jin-Ho
    • Korean Journal of Veterinary Research
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    • v.51 no.2
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    • pp.107-115
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    • 2011
  • Theileria (T.) buffeli (formerly T. sergenti/T. orientalis) is the major hemo-protozoan distributed in the Far East Asian countries such as Korea, China and Japan. It is responsible for the clinical symptoms of anorexia, ateliosis, anemia, fever and icterus. It also causes abortion and sudden death under severe cases, resulting in economic losses for many livestock farms. The objective of this study was to analyze the genetic diversity of the major surface protein (Msp) gene in T. buffeli in Holstein in Korea, and we characterized the association of the diversification of the Msp gene and its relationship with the pathogenicity of Theileria. For this, complete blood counts and Theileria PCR sequence analysis were performed from 57 Holstein in Jeju Island. A total of 26 PCR positive Holstein (16 anemic and 10 non-anemic) were then randomly selected based on 18s rRNA sequence typing of the Theileria Msp gene. The DNA sequence of the T. buffeli Msp gene in Holstein showed 99.0%, 99.2%, 99.9%, 99.5%, 98.7%, 98.4% and 98.4% homology with T. sergenti, Theileria spp., T. sergenti, Theileria spp., Theileria spp., Theileria spp. and Theileria spp., respectively. The result showed a genetic variation of 57.7% (type I), 3.8% (type II), 15.4% (type III), 7.7% (type IV), 13.5% (type V) and 1.9% (type VI). Type I is the most frequent type in both anemic and non-anemic Holstein while type II was found in only non-anemic Holstein. This results of our study help confirm the diversity of Msp gene types and demonstrate that the gene type distribution of Msp genes varies among Theileria-infected Holstein in Jeju Island.

Genetic Relatedness within Streptococcus pneumoniae Serotype 19F and 23F Isolates in Korea by Pulsed-Field Gel Electrophoresis

  • Lee, Kwang-Jun;Bae, Song-Mee;Hwang, Kyu-Jam;Lee, Young-Hee;Kim, Ki-Sang
    • Journal of Microbiology
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    • v.41 no.1
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    • pp.1-6
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    • 2003
  • The genetic relatedness of multidrug-resistant pneumococcal isolates of serotypes 19F and 23F was investigated. The DNA fragments digested with Sma I were resolved by pulsed-field gel electrophoresis (PFGE). PFGE analysis of 365. pneumoniae isolates showed 13 different patterns. Among 22 isolates of serotype 19F, 9 different PFGE patterns were present and 14 isolates of serotype 23F isolates represented 5 distinct PFGE patterns. Two isolates of serotype 19F and six isolates of serotype 23F shared the same PFGE pattern (Pattern I). Based on the genetic relatedness within the strains (one genetic cluster was defined as having more than 85% homology), we divided the pneumococcal strains into genefic clusters (Ⅰ, II, III, IV, V, and VI). The 22 strains of serotype 19F belonged to five distinct genetic clusters (I, II, III, IV, V and VI) and 14 strains of serotype 23F represented two genetic clusters (I and II ). These results showed that strains of serotype 19F are genetically more diverse than those of serotype 23F, Serotype 19F isolates with PFGE patterns H and I appeared to be less related to those of the remaining PFCE patterns (A to G) (less than 60% genetic relatedness), but those strains were genetically closely related with serotype 23f. These results suggest that the latter isolates originated from horizontal transfer of the capsular type 19F gene locus to 23F pneumococcal genotypes. In conclusion, the multidrug-resistant pneumococcal isolates of serotype 19f and 23F isolated in Korea are the result of the spread of a limited number of resistant clones.