• Title/Summary/Keyword: $p_n$-sequences

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Purification and Characterization of a Novel Antifungal Protein from Paenibacillus macerans PM1 Antagonistic to Rice Blast Fungus, Pyricularia oryzae

  • Bae, Dong-Won;Kawk, Weon-Sik;Lee, Joon-Taek;Son, Dae-Young;Chun, Sung-Sik;Kim, Hee-Kyu
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.805-810
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    • 2000
  • An antifungal protein antagonistic to the rice blast fungus, Pyricularia oryzae was purified from Paenibacillus macerans PM-1 by ammonium sulfate fractionation, Q Sepharose Fast Flow column chromatography, Phenyl Sepharose CL-4B column chromatography and Superose 12 gen filtration. An apparent molecular mass of the purified antifungal protein was determined as 8 kDa by SDS-PAGE and 9 kDa by analytical gel filtration, respectively, suggesting that the purified protein is a monomer. The antifungal protein was stable at pH range from 7-12 and up to $100^{\circ}C$. The protein was also stable at 0.1-1% Tween 20 and Triton X-100. The N-terminal amino acid sequence of the antifungal protein was Thr-Glu-Leu-Pro-Leu-Gly-Ile-Val-Met-Asp-Lys-Tyr-Thr-Asp-Ala-Phe-Lys-Phe-Asp-Met-Phe. Comparison of the determined sequence with other peptide and DNA sequences did not reveal homology at all. Therefore, the purified antifungal protein was speculated to be a novel protein. The condidial germination in vitro of P. oryzae KJ301:93-39 by the purified protein ($5.9{\mu} g/ml$) was limited to $9{\pm}3.2%$ only, compared with $69{\pm}2.4%$ of the control. Ungerminated conidia were swollen at basa and mid cell by the purified protein. In vivo bioassay for inhibition of conidial germination of P. oryzae KJ 301, one of the most predominating racesin Korea. the purified protein ($5.9{\mu} g/ml$)strongly inhibited the conidial germination. The conidia, even though germinated, could not develop any further to produce appressoria efficiently.

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Molecular Characterization of Various Trichomonad Species Isolated from Humans and Related Mammals in Indonesia

  • Kamaruddin, Mudyawati;Tokoro, Masaharu;Rahman, Md. Moshiur;Arayama, Shunsuke;Hidayati, Anggi P.N.;Syafruddin, Din;Asih, Puji B.S.;Yoshikawa, Hisao;Kawahara, Ei
    • Parasites, Hosts and Diseases
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    • v.52 no.5
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    • pp.471-478
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    • 2014
  • Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.

Alternative Sigma Factor HrpL of Pectobacterium carotovorum 35 is Important for the Development of Soft-rot Symptoms

  • Nam, Hyo-Song;Park, Ju-Yeon;Kang, Beom-Ryong;Lee, Sung-Hee;Cha, Jae-Soon;Kim, Young-Cheol
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.111-120
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    • 2011
  • A bacterial artificial chromosome library of Pectobacterium carotovorum 35 was constructed to characterize the genome and to sequence its hrp region. The hrp cluster of P. carotovorum 35 consisted of 26 open reading frames in five operons. A promoter-based green fluorescent protein technology was used to identify the genes regulated by the alternative sigma factor, HrpL, in P. carotovorum 35. The majority of the selected clones contained the hrpJ operon promoter sequence, which harbors a hrp box, but no putative hrp boxes were detected within the promoter sequences of two other hrpL-regulated genes encoding for pectate lyase and large repetitive protein. Although the promoters of five other hrp operons also contained hrp boxes, their expression was not HrpL-dependent in the promoter-based selection in E. coli. However, transcriptional analysis showed that expression from all operons harboring hrp boxes, except for the hrpN operon, was reduced significantly in the hrpL mutant. The severity of soft-rot symptoms when the hrpL mutant was applied to the surface of tobacco leaves, mimicking natural infection, was greatly attenuated. These results indicate that the hrpL gene of P. carotovorum 35 may be involved in the development of soft-rot symptoms.

De novo gene set assembly of the transcriptome of diploid, oilseed-crop species Perilla citriodora

  • Kim, Ji-Eun;Choe, Junkyoung;Lee, Woo Kyung;Kim, Sangmi;Lee, Myoung Hee;Kim, Tae-Ho;Jo, Sung-Hwan;Lee, Jeong Hee
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.293-301
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    • 2016
  • High-quality gene sets are necessary for functional research of genes. Although Perilla is a commonly cultivated oil crop and vegetable crop in Southeast Asia, the quality of its available gene set is insufficient. To construct a high-quality Perilla gene set, we sequenced mRNAs extracted from different tissues of Perilla citriodora, the wild species (2n = 20) of Perilla. To make a high-quality gene set for P. citriodora, we compared the quality of assemblies produced by Velvet and Trinity, the two well-known de novo assemblers, and improved the de novo assembly pipeline by optimizing k-mers and removing redundant sequences. We then selected representative transcripts for loci according to several criteria. The improved assembly yielded a total of 86,396 transcripts and 38,413 representative transcripts. We evaluated the assembled transcripts by comparing them to 638 homologous Arabidopsis genes involved in fatty acid and TAG biosynthesis pathways. High proportions of full-length genes and transcripts in the assembled transcripts matched known genes in other species, indicating that the P. citriodora gene set can be applied in future functional studies. Our study provides a reference P. citriodora gene set for further studies. It will serve as valuable genetic resource to elucidate the molecular basis of various metabolisms.

A Parallel Processing System for Visual Media Applications (시각매체를 위한 병렬처리 시스템)

  • Lee, Hyung;Pakr, Jong-Won
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.27 no.1A
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    • pp.80-88
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    • 2002
  • Visual media(image, graphic, and video) processing poses challenge from several perpectives, specifically from the point of view of real-time implementation and scalability. There have been several approaches to obtain speedups to meet the computing demands in multimedia processing ranging from media processors to special purpose implementations. A variety of parallel processing strategies are adopted in these implementations in order to achieve the required speedups. We have investigated a parallel processing system for improving the processing speed o f visual media related applications. The parallel processing system we proposed is similar to a pipelined memory stystem(MAMS). The multi-access memory system is made up of m memory modules and a memory controller to perform parallel memory access with a variety of combinations of 1${\times}$pq, pq${\times}$1, and p${\times}$q subarray, which improves both cost and complexity of control. Facial recognition, Phong shading, and automatic segmentation of moving object in image sequences are some that have been applied to the parallel processing system and resulted in faithful processing speed. This paper describes the parallel processing systems for the speedup and its utilization to three time-consuming applications.

Purification and Characterization of Laccase from Basidiomycete Fomitella fraxinea

  • Park, Kyung-Mi;Park, Sang-Shin
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.670-675
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    • 2008
  • A laccase was isolated from the culture filtrate of the basidiomycete Fomitella fraxinea. The enzyme was purified to electrophoretical homogeneity using ammonium sulfate precipitation, anion-exchange chromatography, and gel-filtration chromatography. The enzyme was identified as a monomeric protein with a molecular mass of 47 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel-filtration chromatography, and had an isoelectric point of 3.8. The N-terminal amino acid sequence for the enzyme was ATXSNXKTLAAD, which had a very low similarity to the sequences previously reported for laccases from other basidiomycetes. The optimum pH and temperature for 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) were 3.0 and $70^{\circ}C$, respectively. The enzyme also showed a much higher level of specific activity for ABTS and 2,6-dimethoxyphenol (DMP), where the $K_m$ values of the enzyme for ABTS and 2,6-DMP were 270 and $426{\mu}M$, respectively, and the $V_{max}$ values were 876 and $433.3{\mu}M/min$, respectively. The laccase activity was completely inhibited by L-cysteine, dithiothreitol (DTT), and sodium azide, significantly inhibited by $Ni^+,\;Mn^{2+}$, and $Ba^{2+}$, and slightly stimulated by $K^+$ and $Ca^{2+}$.

Detection of Co-Infection of Notocactus leninghausii f. cristatus with Six Virus Species in South Korea

  • Park, Chung Hwa;Song, Eun Gyeong;Ryu, Ki Hyun
    • The Plant Pathology Journal
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    • v.34 no.1
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    • pp.65-70
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    • 2018
  • Co-infection with two virus species was previously reported in some cactus plants. Here, we showed that Notocactus leninghausii f. cristatus can be co-infected with six different viruses: cactus mild mottle virus (CMMoV)-Nl, cactus virus X (CVX)-Nl, pitaya virus X (PiVX)-Nl, rattail cactus necrosis-associated virus (RCNaV)-Nl, schlumbergera virus X (SchVX)-Nl, and zygocactus virus X (ZyVX)-Nl. The coat protein sequences of these viruses were compared with those of previously reported viruses. CMMoV-Nl, CVX-Nl, PiVX-Nl, RCNaV-Nl, SchVX-Nl, and ZyVX-Nl showed the greatest nucleotide sequence homology to CMMoV-Kr (99.8% identity, GenBank accession NC_011803), CVX-Jeju (77.5% identity, GenBank accession LC12841), PiVX-P37 (98.4% identity, GenBank accession NC_024458), RCNaV (99.4% identity, GenBank accession NC_016442), SchVX-K11 (95.7% identity, GenBank accession NC_011659), and ZyVX-B1 (97.9% identity, GenBank accession NC_006059), respectively. This study is the first report of co-infection with six virus species in N. leninghausii f. cristatus in South Korea.

Certification of Gibroblase Cell Adhesion and Spreading Mediated by Arg-Gly-Asp (RGD) Sequence on Thermo-Reversible Hydrogel

  • NA, KUN;DONG-WOON KIM;KEUN-HONG PARK
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.922-927
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    • 2001
  • In an effort to regulate the mammalian cell behavior in entrapment with a gel, we have functionalized hydrogels with the putative cell-binding (-Arg-Gly-Asp-)(RGD) domain. An adhesion molecule of Gly-Arg-Gly-Asp-Ser (GRGDS) peptides, a cell recognition ligand, was induced into thermo-reversible hydrogels, composed of N-isopropylacrylamide with small amounts of acrylic acid (typically 2-5 $mol\%$ in feed), as a biomimetic extracellular matrix (ECM). The GRGDS containing a p(NiPAAm-co-AAc) copolymer gel was studied in vitro for its ability to promote the spreading and viability of cells by introducing a GRGDS sequence. Hydrogel with no adhesion molecule was a poor ECM for adhesion, permiting spreading of only $3\%$ of the seeded cells for 36h. By immobilizing the peptide linkage into the hydrogel, the conjugation of RGD promoted $50\%$ of proliferation for 36h. However, the GREDS sequence, nonadhesive peptide linkage, conjugated hydrogel showed only $5\%$ of the seeded cell for the same time period. In addition, with the serum-free medium, only GRGDS peptides conjugated to hydrogel was able to promotecell spreading, while there was no cell proliferation in the hydrogel without GRGDS. Thus, the GRGDS peptide-conjugated thermo-reversible hydrogel specifically mediated the cell spreading. This result suggests that utilization of peptide sequences conjugating with the cell-adhesive motifs can enhance the degree of cell surface interaction and influence the long-term formation of ECM in vitro.

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Mechanism of Action of Anticancer Drug Aziridinylbenzoquinones: Involvement of DT-diaphorase (DNA에 결합하는 항암제의 작용기전)

  • Lee, Chong-Soon-
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.11a
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    • pp.147-172
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    • 1994
  • Aziridinylbenzoquinones such as 3, 6-diaziridinyl-1, 4-benzoquinone (DZQ) and its 2, 5-methyl analog (MeDZQ) require bioreductive activation in order to elicit their anticancer activities. To determine the involvement of DTD in the activation of these drugs, we have used a ligation-mediated polymerase chain reaction to map the intracellular alkylation sites in a sing1e copy gene at the nucleotide level. We have performed this analysis in two human colon carcinoma cells, one proficient (HT-29) and one deficient (BE) in DT-diaphorase (DTD) activity. In the DTD proficient HT-29 cell line, DZQ and MeDZQ were found to alkylate both 5'-(A/T)G(C)-3' and 5'-(A/T)A-3' sequences. This is consistent with the nucleotide preferences observed when DZQ and MeDZQ are activated by purified DTD to reactive metabolites capable of alkylating DNA in vitro [Lee, C. -S., Hartley, J. A., Berardini, M. D., Butler, J., Siegel., D., Ross, D., & Gibson, N. W. (1992) Biochemistry, 31: 3019-3025]. Surprisingly in the DTD-deficient BE cell line a pattern of alkylation induced by DZQ and MeDZQ similar to that observed in the DTD-proficient HT-29 cells was observed. This suggests that reductive enzymes other than DTD can be involved in activating DZQ and MeDZQ to DNA reactive species in vivo.

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Genetic Diversity of 14 Indigenous Grey Goose Breeds in China Based on Microsatellite Markers

  • Tu, Yunjie;Chen, K.W.;Zhang, S.J.;Tang, Q.P.;Gao, Y.S.;Yang, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.1
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    • pp.1-6
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    • 2006
  • This experiment first cloned some microsatellite sequences for goose species by magnetic beads enriched method and studied the genetic structure research of 14 indigenous grey goose breeds using 19 developed and 12 searched microsatellite markers with middle polymorphism. According to the allele frequencies of 31 microsatellite sites, mean heterozygosity (H), polymorphism information content (PIC) and $D_A$ genetic distances were calculated for 31-microsatellite sites. The results showed that 25 of 31microsatellite sites were middle polymorphic, so the 25 microsatellite markers were effective markers for analysis of genetic relationship among goose breeds. The mean heterozygosity was between 0.4985 and 0.6916. The highest was in the Xupu (0.6916), and in the Yan was the lowest (0.4985) which was consistent with that of PIC. The phylogenetic tree was completed through analysis of UPGMA. Fencheng Grey, Shoutou, Yangjiang and Magang were grouped firstly, then Xongguo Grey, Wugang Tong, Changle and Youjiang were the second group; Gang, Yan Xupu and Yili were the third group; Yongkang Grey and Wuzeng were the fourth group. The results could provide basic molecular data for the research on the characteristics of local breeds in the eastern China, and a scientific basis for the conservation and utilization of those breeds.