• Animal cells and systems
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• v.11 no.1
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• pp.39-50
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• 2007

Tau plays a role in numerous neuronal processes, such as vesicle transport, microtubule-plasma membrane interaction and intracellular localization of proteins. SUMO (Small Ubiquitin-like Modifier) modification (SUMOylation) appears to regulate diverse cellular processes including nuclear transport, signal transduction, apoptosis, autophagy, cell cycle control, ubiquitin-dependent degradation, as well as gene transcription. We noticed that putative SUMOylation site is localized at $^{340}K$ of $Tau(^{339}VKSE^{342})$ with the consensus sequence information (${\Phi}KxE$ ; where ${\Phi}$ represents L, I, V or F and x is any amino acid). In this report, we demonstrated that $^{340}K$ of Tau is the SUMOylation site and that a point mutant of Tau S214E (an analog of the phospho $^{214}S$ Tau) promotes its SUMOylation at $^{340}K$ and its nuclear or nuclear vicinity localization, by co-immunoprecipitation and confocal microscopy analysis. Further, we demonstrate that the Tau S214E (neither Tau S214A nor Tau K340R) mutant increases its protein stability. However, the SUMOylation at $^{340}K$ of Tau did not influence cell survival, as determined by FACS analysis. Therefore, our results suggested that the phosphorylation of Tau on $^{214}S$ residue promotes its SUMOylation on $^{340}K$ residue and nuclear vicinity localization, and increases its stability, without influencing cell survival.

• The Plant Pathology Journal
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• v.37 no.1
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• pp.36-46
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• 2021

Acidovorax citrulli (Ac) is the causal agent of bacterial fruit blotch (BFB) in watermelon, a disease that poses a serious threat to watermelon production. Because of the lack of resistant cultivars against BFB, virulence factors or mechanisms need to be elucidated to control the disease. Glycerol-3-phosphate dehydrogenase is the enzyme involved in glycerol production from glucose during glycolysis. In this study, we report the functions of a putative glycerol-3-phosphate dehydrogenase in Ac (GlpdAc) using comparative proteomic analysis and phenotypic observation. A glpdAc knockout mutant, AcΔglpdAc(EV), lost virulence against watermelon in two pathogenicity tests. The putative 3D structure and amino acid sequence of GlpdAc showed high similarity with glycerol-3-phosphate dehydrogenases from other bacteria. Comparative proteomic analysis revealed that many proteins related to various metabolic pathways, including carbohydrate metabolism, were affected by GlpdAc. Although AcΔglpdAc(EV) could not use glucose as a sole carbon source, it showed growth in the presence of glycerol, indicating that GlpdAc is involved in glycolysis. AcΔglpdAc(EV) also displayed higher cell-to-cell aggregation than the wild-type bacteria, and tolerance to osmotic stress and ciprofloxacin was reduced and enhanced in the mutant, respectively. These results indicate that GlpdAc is involved in glycerol metabolism and other mechanisms, including virulence, demonstrating that the protein has pleiotropic effects. Our study expands the understanding of the functions of proteins associated with virulence in Ac.

• Animal Bioscience
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• v.37 no.2_spc
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• pp.396-403
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• 2024

Objective: Monofluoroacetate (MFA) is a potent toxin that blocks ATP production via the Krebs cycle and causes acute toxicity in ruminants consuming MFA-containing plants. The rumen bacterium, Cloacibacillus porcorum strain MFA1 belongs to the phylum Synergistota and can produce fluoride and acetate from MFA as the end-products of dehalorespiration. The aim of this study was to identify the genomic basis for the metabolism of MFA by this bacterium. Methods: A draft genome sequence for C. porcorum strain MFA1 was assembled and quantitative transcriptomic analysis was performed thus highlighting a candidate operon encoding four proteins that are responsible for the carbon-fluorine bond cleavage. Comparative genome analysis of this operon was undertaken with three other species of closely related Synergistota bacteria. Results: Two of the genes in this operon are related to the substrate-binding components of the glycine reductase protein B (GrdB) complex. Glycine shares a similar structure to MFA suggesting a role for these proteins in binding MFA. The remaining two genes in the operon, an antiporter family protein and an oxidoreductase belonging to the radical S-adenosyl methionine superfamily, are hypothesised to transport and activate the GrdB-like protein respectively. Similar operons were identified in a small number of other Synergistota bacteria including type strains of Cloacibacillus porcorum, C. evryensis, and Pyramidobacter piscolens, suggesting lateral transfer of the operon as these genera belong to separate families. We confirmed that all three species can degrade MFA, however, substrate degradation in P. piscolens was notably reduced compared to Cloacibacillus isolates possibly reflecting the loss of the oxidoreductase and antiporter in the P. piscolens operon. Conclusion: Identification of this unusual anaerobic fluoroacetate metabolism extends the known substrates for dehalorespiration and indicates the potential for substrate plasticity in amino acid-reducing enzymes to include xenobiotics.

• Journal of Animal Science and Technology
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• v.54 no.4
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• pp.241-246
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• 2012

The goal of this study was to verify the phylogenetic status of the Korean native goats (KNG). We determined the complete sequence of the mitochondrial cytochrome b gene in 48 goats among four populations. We also analyzed genetic variability within goats, and a phylogenetic analysis was performed by comparison with other country's goats. Three nucleotide substitutions were detected, and two of these were missense mutations that occurred due to a substitution of amino acid. Four haplotypes were defined from KNG. Three of these haplotypes were only found in the Chinese goat. However, the other haplotype was KNG-specific. In the phylogenetic analysis, four clades (A~D) were classified among domestic goats, and the KNG was classified into clade 1 that estimated as lineage A based on the D-loop sequence. Each haplotype from the KNG was clustered closely with that of the Chinese goat. The results of haplotype distribution and phylogenetic location suggest that strong gene flow occurred from China to the Korean Peninsula.

• Journal of Life Science
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• v.26 no.4
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• pp.419-430
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• 2016

Powdery mildew disease caused by Podosphaera xanthii is a major concern for Cucumis melo production worldwide. Knowledge on genetic behavior of the related genes and their modulating phytohormones often offer the most efficient approach to develop resistance against different diseases. Mildew Resistance Locus O (MLO) genes encode proteins with seven transmembrane domains that have significant function in plant resistance to powdery mildew fungus. We collected 14 MLO genes from ‘Melonomics’ database. Multiple sequence analysis of MLO proteins revealed the existence of both evolutionary conserved cysteine and proline residues. Moreover, natural genetic variation in conserved amino acids and their replacement by other amino acids are also observed. Real-time quantitative PCR expression analysis was conducted for the leaf samples of P. xanthii infected and phyto-hormones (methyl jasmonate and salicylic acid) treated plants in melon ‘SCNU1154’ line. Upon P. xanthii infection using 7 different races, the melon line showed variable disease reactions with respect to spread of infection symptoms and disease severity. Three out of 14 CmMLO genes were up-regulated and 7 were down-regulated in leaf samples in response to all races. The up- or down-regulation of the other 4 CmMLO genes was race-specific. The expression of 14 CmMLO genes under methyl jasmonate and salicylic acid application was also variable. Eleven CmMLO genes were up-regulated under salicylic acid treatment, and 7 were up-regulated under methyl jasmonate treatments in C. melo L. Taken together, these stress-responsive CmMLO genes might be useful resources for the development of powdery mildew disease resistant C. melo L.

• International Journal of Industrial Entomology and Biomaterials
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• v.18 no.1
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• pp.18-27
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• 2009

B. thuringiensis strain LY-99 belonging to subsp. alesti (H3a3c), was isolated from Chinese tobacco warehouse and showed significantly high toxicity to Plutella xylostella. For the identification of the cry1-type genes from B. thuringiensis LY-99, an extended multiplex PCRrestriction fragment length polymorphism (PCRRFLP) method was established by using two pairs of universal primers based on the conserved regions of the cry1-type genes to amplify around 2.4 kb cry1-type gene fragments. Then the DNA fragment was cloned into pGEM-T Easy vector and digested with EcoRI and EcoRV enzymes. Through this method, a known cry1-type gene was successfully identified from the reference strain, B. thuringiensis subsp. alesti. In addition, the RFLP patterns revealed that B. thuringiensis LY-99 included a novel cry1A-type gene in addition to cry1Aa, cry1Ac, cry1Be and cry1Ea genes. The novel cry1A-type gene was designated cry1Ah2 (Genbank accession No DQ269474). An inverse PCR method was used to amplify the flank regions of cry1Ah2 gene. Finally, 3143 bp HindIII fragment from B. thuringiensis LY-99 plasmid DNA including 5' region and partial ORF was amplified, and sequence analysis revealed that cry1Ah2 gene from LY-99 showed 89.31% of maximum sequence similarity with cry1Ac1 crystal protein gene. In addition, the deduced amino acid sequence of Cry1Ah2 protein shared 87.80% of maximum identity with that of Cry1Ac2. This protein therefore belongs to a new class of B. thuringiensis crystal proteins.

• Korean Journal of Microbiology
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• v.35 no.3
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• pp.205-210
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• 1999

16brio cholerne is an important pathogenic organism that causes dimhea in human beings. V ciaoleroe KNIH002 was isolated from patients suffering with dian.heal disease in Korea. From Southern hybridization using the amplified PCR product of 307 bp as a probe. which was obtained from PCR reaction using primer detecting cholera toxin gene, we have found that the c b gene located in 4.5-kb fragmenl double digested with Pstl and BgllI of the chromosome. Therefore, we made mini-libraries of the isolate using PstI and Bgm restriction endonuclease and pBluescript SKU(+) vector. As a result. we cloned 4.5-kb PstI-BglII fragment containing the c a gene encoding a cholera toxin from the constructed mini-libraries of V olzolerae KNlH002 by colony hybridization using the same probes. This recombinant plasmid was named pCTX75. E. coii XL1- Blue harboring pCTX75 showed the cytotoxicity on Chinese Hamster Ovary cells. From the sequencing of he cloned recombinant plasmid, we confinned that it has virulence gene cassette consisting of ace, zot, ctx.4 and cf"~B gene. The ace and zot genes were composed of 291 hp and 1.200 bp with ATG initiation codon and TGA lennination codon, respectively. Nucleotide sequence of the ace gene exhibited 100% identity with that of V cholera E7946 El Tor Ogawa strains. But, nucleolide and amino acid sequence comparison of the zot gene exhibited 99% and 98.8% identity with that of V cholerae 395 Classical Ogawa stram, respectively. Specially. the Ala-100, Ala-272 and Ala-281 sites of Zoi polypeptide presented in V choleme 395 Classical Ogawa strain are replaced by Val in V cholerae KNIH002.

• The KIPS Transactions:PartD
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• v.15D no.2
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• pp.155-162
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• 2008

Biological sequences such as DNA sequences and amino acid sequences typically contain a large number of items. They have contiguous sequences that ordinarily consist of hundreds of frequent items. In biological sequences analysis(BSA), a frequent contiguous sequence search is one of the most important operations. Many studies have been done for mining sequential patterns efficiently. Most of the existing methods for mining sequential patterns are based on the Apriori algorithm. In particular, the prefixSpan algorithm is one of the most efficient sequential pattern mining schemes based on the Apriori algorithm. However, since the algorithm expands the sequential patterns from frequent patterns with length-1, it is not suitable for biological dataset with long frequent contiguous sequences. In recent years, the MacosVSpan algorithm was proposed based on the idea of the prefixSpan algorithm to significantly reduce its recursive process. However, the algorithm is still inefficient for mining frequent contiguous sequences from long biological data sequences. In this paper, we propose an efficient method to mine maximal frequent contiguous sequences in large biological data sequences by constructing the spanning tree with the fixed length. To verify the superiority of the proposed method, we perform experiments in various environments. As the result, the experiments show that the proposed method is much more efficient than MacosVSpan in terms of retrieval performance.

• Korean journal of applied entomology
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• v.36 no.4
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• pp.331-340
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• 1997

Hyalophora cecropia attacin-like antibacterial gene was isolated from Bombyx mori induced with nonpathogenic bacteria. It was expressed in Spodopfera frugiperda 9 (Sf9 cells using baculovirus expression vector system (BEVS), and examined its antibacterial activity. With a cDNA library constructed from fifthinstar B. mori injected with Escherichia coli(4 X IOhcellsllarva), differential screening was performed using naive and induced mRNA probes. BmInc6 clone was screened by partial nucleotide sequence and GenBank database analysis. A complete nucleotide sequence of Bmlnc6 cDNA was determined (GenBank, AF005384). Its insert size was 852 bp and had open reading frame that started translation at position 35 and stopped at 679. And its putative polyadenylational signal existed at 812 bp. The number of amino acid deduced from Bmlnc6 cDNA was 214 and hydropathy analysis showed that this peptide was hydrophilic. This peptide deduced by BmInc6 was named nuecin. When the nuecin gene was expressed in Sf9 cells using BEVS, about 950 bp of the transcripts was detected. In addition, SDS-PAGE analysis showed that the molecular weights of intracellular expressed protein and the mature protein secreted to culture media were approximately 23 and 20 kDa, respectively. The antibacterial activity of nuecin against E. coli and Bacillus subtilis was significantly high, demonstrating that nuecin had a wider antibacterial spectrum with gram negative and positive bacteria than attacin.

• Research in Plant Disease
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• v.24 no.4
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• pp.285-291
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• 2018

Two isolates of Strawberry mild yellow edge virus were newly isolated in strawberry (Fragaria x ananassa) cultivar Selhyang and Kamhong from Korea. The complete nucleotide sequence of the coat protein (CP) of two Korean isolates were determined and analyzed. Sequence identity of nucleotide and amino acid between SH and KH isolates was 90.4% and 95.5%, respectively. The comparison of three Korean isolates including previously reported KNS1 with 45 SMYEV sequences from other countries deposited in GenBank database revealed an identity ranging from 81.2% to 100%. The phylogenetic analysis of CP of all SMYEV isolates showed the five subgroups (I-V), with Korean isolates being divided into two different subgroups. The isolates KH and KNS1 were included in subgroup I, whereas SH was included in subgroup IV which is new phylogenetic subgroup. Genetic diversity analysis indicated that new subgroup had greater variability and nucleotide diversity between subgroups resulted in values ranging from 0.0863 to 0.18004. This report represents the first molecular characterization of SMYEV isolates from Korea.