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

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Sensitivity of Pseudomonas syringae to Bovine Lactoferrin Hydrolysates and Identification of a Novel Inhibitory Peptide

  • Kim, Woan-Sub;Kim, Pyeung-Hyeun;Shimazaki, Kei-ichi
    • Food Science of Animal Resources
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    • v.36 no.4
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    • pp.487-493
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    • 2016
  • The antimicrobial activity of bovine lactoferrin hydrolysates (bLFH) was measured against Pseudomonas strains (P. syringae and P. fluorescens) in vitro. To compare susceptibility to bLFH, minimal inhibitory concentration (MIC) values were determined using chemiluminescence assays and paper disc plate assays. Antimicrobial effect against P. fluorescens was not observed by either assay, suggesting that bLFH did not exhibit antimicrobial activity against P. fluorescens. However, a significant inhibition of P. syringae growth was observed in the presence of bLFH. The addition of bLFH in liquid or solid medium inhibited growth of P. syringae in a dose-dependent manner. Furthermore, a bLFH peptide with antimicrobial activity toward P. syringae was isolated and identified. The N-terminal amino acid sequences of thus obtained antimicrobial bLFH peptides were analyzed by a protein sequencer and were found to be Leu-Arg-Ile-Pro-Ser-Lys-Val-Asp-Ser-Ala and Phe-Lys-Cys-Arg-Arg-Trp-Gln-Trp-Arg-Met. The latter peptide sequence is known to be characteristic of lactoferricin. Therefore, in the present study, we identified a new antimicrobial peptide against P. syringae, present within the N-terminus and possessing the amino acid sequence of Leu-Arg-Ile-Pro-Ser-Lys-Val-Asp-Ser-Ala.

Distribution of Telomeric DNA in Korean Native Chicken Chromosomes (한국 재래닭 염색체의 텔로미어 분포 양상)

  • Sohn, Sea-Hwan;Cho, Eun-Jeong
    • Korean Journal of Poultry Science
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    • v.37 no.3
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    • pp.247-253
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    • 2010
  • Telomeres are nucleoprotein structures at the ends of chromosomes consisting of DNA sequences arranged in tandemly repeated units $(TTAGGG)_n$. However, this hexamers can also occur at non-telomeric sites in some avians and vertbrate. This study was carried out to present the distribution of telomeric DNA sequences in Korean Native Chicken chromosomes. The fluorescence in situ hybridization technique using a telomeric DNA probe was performed to the metaphase spreads of chicken early embryonic cells. Telomeric DNA signals were detected at the ends of chromosomes including macrochromosomes and microchromosomes. In chicken, surprisingly, chromosome 1 showed very distinct interstitial telomeric DNA hybridization patterns which located two interstitial sites in the p-arm at 1p11 and 1p23, and one in the q-arm at 1q32. In chromosome number 2 and 3 also displayed interstitial telomeric signals (ITS) in the long arms at 2q24 and 3q32, respectively. The pattern of telomeric DNA distribution in Korean Native Chicken chromosomes was in agreement with a previously reported in Gallus domesticus. The relative amount of telomeric DNA sequences in each macrochromosomes ranged from 4.6% to 16.3%. Distribution of telomeric DNAs at the end of p-arm was much more than that of q-arm in almost chicken chromosomes. The distribution of ITS in chicken chromsomes implicate to tandem chromosome fusions that might have occurred during the process of karyotype evolution.

Rapid Determining for Subtypes and Pandemic Type of Swine Influenza Virus by Diagnostic One-step RT-PCR (진단용 one-step RT-PCR을 통한 돼지 인플루엔자 바이러스의 아형 및 pandemic 유형에 대 한 신속한 결정)

  • Kim, Gwang Il;Kim, Jee In;Kwon, Jin-Hyeap;Min, Yoo Hong;Kang, Joo Il;Lee, Chang-Ho;Kim, Sung-Hee;Lim, Jae-Hwan
    • Journal of Life Science
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    • v.28 no.5
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    • pp.555-562
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    • 2018
  • Swine influenza virus (SIV) causes one of the most common diseases of the pig population, and its subtypes are determined by hemagglutinin (HA) and neuraminidase (NA). Recently, the SIV subtype diagnosis has been developed. The method using antigen-antibody reaction rather than PCR was mainly used because of the large change in the ribonucleotide sequences of SIV. Here, we have developed 10 diagnostic primer sets through multi-nucleotide sequences alignment of spreaded SIV since 2008 in Korea and then optimized the reaction of the one-step RT-PCR for rapid determination of SIV subtype. In addition, specific primers were designed to early determine the pandemic SIV by detecting unique M sequences proven in highly infectious and virulent subtypes of the influenza H1N1 (pH1N1). Here, some of the SIVs spread in Korea from 2008 to 2014 have been tested to determine the subtypes and pandemic potential of SIV. All diagnostic primer sets were found to be able to accurately determine the SIV subtype and to detect the pandemic SIV. In conclusion, it was confirmed that the optimized one-step RT-PCR analysis using these primer sets is useful for rapid diagnosis of SIV subtypes. These results can be used for development of SIV subtype diagnostic kit to early detect before virulent SIV spreads do.

Development of a novel endolysin, PanLys.1, for the specific inhibition of Peptostreptococcus anaerobius

  • Joonbeom Moon;Hanbeen Kim;Dongseok Lee;Jakyeom Seo
    • Animal Bioscience
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    • v.36 no.8
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    • pp.1285-1292
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    • 2023
  • Objective: The objective of this study was to develop a novel endolysin (PanLys.1) for the specific killing of the ruminal hyper-ammonia-producing bacterium Peptostreptococcus anaerobius (P. anaerobius). Methods: Whole genome sequences of P. anaerobius strains and related bacteriophages were collected from the National Center for Biotechnology Information database, and the candidate gene for PanLys.1 was isolated based on amino acid sequences and conserved domain database (CDD) analysis. The gene was overexpressed using a pET system in Escherichia coli BL21 (DE3). The lytic activity of PanLys.1 was evaluated under various conditions (dosage, pH, temperature, NaCl, and metal ions) to determine the optimal lytic activity conditions. Finally, the killing activity of PanLys.1 against P. anaerobius was confirmed using an in vitro rumen fermentation system. Results: CDD analysis showed that PanLys.1 has a modular design with a catalytic domain, amidase-2, at the N-terminal, and a cell wall binding domain, from the CW-7 superfamily, at the C-terminal. The lytic activity of PanLys.1 against P. anaerobius was the highest at pH 8.0 (p<0.05) and was maintained at 37℃ to 45℃, and 0 to 250 mM NaCl. The activity of PanLys.1 significantly decreased (p<0.05) after Mn2+ or Zn2+ treatment. The relative abundance of P. anaerobius did not decrease after administration PanLys.1 under in vitro rumen conditions. Conclusion: The application of PanLys.1 to modulate P. anaerobius in the rumen might not be feasible because its lytic activity was not observed in in vitro rumen system.

CYCLIC CODES FROM THE FIRST CLASS TWO-PRIME WHITEMAN'S GENERALIZED CYCLOTOMIC SEQUENCE WITH ORDER 6

  • Kewat, Pramod Kumar;Kumari, Priti
    • Bulletin of the Korean Mathematical Society
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    • v.56 no.2
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    • pp.285-301
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    • 2019
  • Let $p_1$ and $p_2$ be two distinct odd primes with gcd($p_1-1$, $p_2-1$) = 6. In this paper, we compute the linear complexity of the first class two-prime Whiteman's generalized cyclotomic sequence (WGCS-I) of order d = 6. Our results show that their linear complexity is quite good. So, the sequence can be used in many domains such as cryptography and coding theory. This article enrich a method to construct several classes of cyclic codes over GF(q) with length $n=p_1p_2$ using the two-prime WGCS-I of order 6. We also obtain the lower bounds on the minimum distance of these cyclic codes.

Analysis of antigenic domain of GST fused major surface protein (p30) fragments of Toxoplasma gondii (융합단백질로 발현된 톡소포자충의 주요막단백질(p30) 절편의 항원성)

  • 남호우;임경심
    • Parasites, Hosts and Diseases
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    • v.34 no.2
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    • pp.135-142
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    • 1996
  • Antigenic domain of jai or surface protein (p30) of Toxoplosmc Sondii was analyzed after polymerase chain reaction (PCR) of its gene fragments. Hydrophilic or hydrophobic moiety of amino acid sequences were expressed as glutathione S-transferase (G57) fusion proteins. Fragments of p30 gene were as follows: 737, total p30 open reading frame (ORF) ; S28, total ORF excluding N-terminal signal sequence and C-terminal hydrophobic sequence; Al9, N-terminal 2/3 parts of A28; A19, N-terminal 2/3 of S28; P9, C-terminal 2/3 part of S28; Z9. middle 1/3 of S28; and 29, C-terminal 1/3 of S28. respectively. Primer of each fragment was synthesized to include clamp sequence of EcoR I restriction site. PCR amplified DNA was inserted info GST (26 kDa) expression vector, PGEX-47-1 to transform into Escheri,hia coei (.JM105 strain). G57 fusion proteins were expressed with IPTG induction as 63. 54, 45, 45, 35, 36. and 35 kDa proteins measured by SDS-PAGE. Each fusion protein was confirmed with G57 detection kit. Western blot analysis with the serum of a toxoplasmosis patient revealed antigenicity in proteins expressed by T37. S28, and Al9 but not those by Pl8. X9, Y10, and Z9. Antigenicity of p30 seems to be located either in N-terminal 115 part in the presence of middle 1/3 part or in the oligopeptides between margins of the first and second 1/3 parts.

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Identification of New Microsatellite Markers in Panax ginseng

  • Kim, Joonki;Jo, Beom Ho;Lee, Kyoung Lyong;Yoon, Eui-Soo;Ryu, Gi Hyung;Chung, Ki Wha
    • Molecules and Cells
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    • v.24 no.1
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    • pp.60-68
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    • 2007
  • Microsatellites, also called simple sequence repeats (SSR), are very useful molecular genetic markers commonly used in crop breeding, species identification and linkage analysis. In the present study, we constructed a microsatellite-enriched genomic library of Panax ginseng, and identified 251 novel microsatellite sequences. Tri-nt repeat units were the most abundant (46.6%), followed by di-nt repeats (35.5%). The $(AG)_n$ motif was most common (23.1%), followed by the $(AAC)_n$ motif (22.3%). From the genotyping of 94 microsatellites using marker-specific primer sets, we identified 11 intraspecific polymorphic markers as well as 14 possible interspecific polymorphic markers differing between P. ginseng and P. quinquefolius. The exact allele structures of the polymorphic markers were determined and the alleles were named. This study represents the first report of the bulk isolation of microsatellites by screening a microsatellite-enriched genomic library in P. ginseng. The microsatellite markers could be useful for linkage analysis, genetic breeding and authentication of Panax species.

Base Specificity for DNA Interstrand Cross-Linking Induced by Anticancer Agent Bizelesin

  • Lee, Chong-Soon;Myung, Pyung-Keun;Gibson, Neil W.
    • Archives of Pharmacal Research
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    • v.19 no.3
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    • pp.191-196
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    • 1996
  • Bizelesin is a promising novel anticancer agent which is known to alkylate N3 of adenine to induce DNA interstrand cross-links (ISC) with in $5^I-TAATTA\; and\; 5^I-TAAAAAA$. We have investigated the base specificity for DNA ISC induced by bizelesin using oligomers containing the cross-linkable sequence $5^I-TAATTA\; and\; 5^I-TAAAAAA$. in which "N" was either A, C, G, or T. An analysis of denaturing polyacrylamide gel showed that bizelesin is able to induce DNA ISC in the duplex oligomer containing sequences $5^I-TAATTA\; and\; 5^I-TAAAAAA$. The formation of interstrand crosslinking did not occur in the sequences $5^I-TAATTA\; and\; 5^I-TAAAAAA$. DNA strand cleavage assay to determine the cross-linking site within $5^I-TAATTA$sequence showed that bizelesin alkylates guanine. These results demonstrate that bizelesin is able to induce DNA ISC at guanine but not at cytosine or thymine. In addition, guanine adducts have been found to be susceptible to DNA strand cleavage by exposure to hot piperidine. The extent of DNA strand cleavage, however, was not 100% efficient in either neutral pH buffer or hot piperidine.

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Molecular Cloning and Nucleotide Sequence of the Gene Encoding Fusion(F) Protein of the Thermostable Newcastle Disease Virus Isolated from a Diseased Pheasant (꿩에서 분리된 Newcastle Disease Virus 내열성주 (CBP)의 Fusion(F) 유전자 클론닝과 염기서열 분석)

  • Chang, Kyung-Soo;Jun, Moo-Hyung;Song, Hee-Jong;Kim, Kui-Hyun;Park, Jong-Hyeon
    • The Journal of Korean Society of Virology
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    • v.28 no.3
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    • pp.233-245
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    • 1998
  • The gene encoding F protein of CBP-1 strain, a heat-stable Newcastle disease virus (NDV) isolated from the diseased pheasants in Korea, was characterized by reverse transcription-polymerase chain reaction (RT-PCR), nucleotide and amino acid sequences. Virus RNA was prepared from the chorioallatoic fluid infected with NDV CBP-1 virus and cDNA was amplified by RT-PCR, cloned and sequenced to analyze. The PCR was sensitive as to detect the virus titer above $2^5$ hemagglutination unit. 1.7kb (1,707bp) size of the cDNA was amplified and cloned into BamHI site of pVL1393 Baculo transfer vector. The nucleotide sequences for F protein were determined by dye terminator cyclic sequencing using four pairs of primers, and 553 amino acid sequences were predicted. In comparison of the nucleotide sequence of F gene of CBP-1 with those of other NDV strains, the homology revealed 88.8%, 98.5% and 98.7% with Kyojungwon (KJW), Texas GB and Beaudette C strains, respectively. As the deduced 553 amino acid sequences of F protein of CBP-1 were compared with those of other NDV strains, the homology appeared 89.9%, 98.7% and 98.9% with KJW, Texas GB and Beaudette C strains, respectively. The putative protease cleavage site (112-116) was R-R-Q-K-R, indicating that CBP-1 strain is velogenic type. The amino acid sequences include 6 sites of N-asparagine-linked glycosylation and 13 cysteine residues. These data indicate that the genotype of CBP-1 strain is more closely associated with the strains of Texas GB and Beaudette C than KJW strain.

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Additional mitochondrial DNA sequences from the dung beetle, Copris tripartitus (Coleoptera: Scarabaeidae), an endangered species in South Korea

  • Hwang, Eun Ju;Jeong, Su Yeon;Wang, Ah Rha;Kim, Min Jee;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.36 no.2
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    • pp.31-41
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    • 2018
  • The dung beetle, Copris tripartitus (Coleoptera: Scarabaeidae), is an endangered insect in South Korea. Previously, partial mitochondrial COI and CytB gene sequences have been used to infer genetic diversity and gene flow of this species in South Korea. In this study, we additionally collected C. tripartitus (n = 35) from one previous locality and two new localities, sequenced COI and CytB genes, and combined these with preexisting data for population genetic analysis. Sequence divergence of current samples showed slightly lower values [4.86% (32 bp) for COI and 4.16% (18 bp) for CytB] than that in the previous study. Nucleotide diversity (${\pi}$) ranged from 0.005336 (Gulupdo) to 0.020756 (Seogwi-dong) in COI and 0.009060 (Aewol-eup) to 0.017464 (Seogwi-dong) in CytB. Seogwi-dong samples that showed the highest ${\pi}$ in the previous study also showed the highest ${\pi}$ in this study for both gene sequences. The newly investigated Gulupdo samples had the lowest haplotype diversity for both gene sequences. They also had the lowest ${\pi}$ for COI and the second lowest ${\pi}$ for CytB. On the other hand, the newly added Haean-dong sample had relatively higher diversity estimates. Gene flow among populations was high, although significant difference was only detected between Gulupdo and Anmado or between Gulupdo and Seogwi-dong for COI sequences (P < 0.05). Considering the high genetic diversity and gene flow in C. tripartitus populations, one major issue regarding conservation seems not to be recovery of genetic diversity.