• Title/Summary/Keyword: sequence homology

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Prevalence study of bovine viral diarrhea virus (BVDV) from cattle farms in Gyeongsangnam-do, South Korea in 2021 (2021년 경남지역 소바이러스성설사 바이러스(BVDV) 감염실태 조사)

  • Son, Yongwoo;Cho, Seonghee;Ji, Jeong-Min;Cho, Jae-Kyu;Bang, Sang-Young;Choi, Yu-Jeong;Kim, Cheol-Ho;Kim, Woo Hyun
    • Korean Journal of Veterinary Service
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    • v.45 no.3
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    • pp.211-219
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    • 2022
  • Bovine viral diarrhea (BVD) is one of the problematic wasting diseases in cattle leading to huge economic losses. This study was conducted to investigate the prevalence of BVD including transient and persistent infection from cattle farms in Gyeongsangnam-do. A total of 2,667 blood samples from 24 farms were collected and the sera were subjected to ELISA to detect BVD virus (BVDV) antigen, Erns. 5' untranslated region (5'-UTR) of BVDV-positive samples was sequenced to identify the genotype, and compared with isolates previously reported elsewhere. There were fourteen BVDV-positive calves from 2,667 samples (positive rate: 0.52%) from first ELISA testing followed by eight persistently infected out of eleven BVDV-positive samples (72.73%) in secondary ELISA that was conducted in at least four weeks suggesting the circulation of BVDV in the area. Sequencing analysis exhibited that thirteen BVDV-positive samples were identified as BVDV-1b and one sample was BVDV-2a. Phylogenetic analysis revealed that the BVDV-1b-positive samples showed the highest homology in nucleotide sequence to Korean isolates collected from Sancheong, Gyeongsangnam-do, while the BVDV-2a-positive sample (21GN7) was more similar to reference strains collected outside South Korea. This study will provide the recent fundamental data on BVD prevalence in Gyeongsangnam-do to be referred in developing strategies to prevent BVDV in South Korea.

Phylogenetic analysis and biological characterization of Stemphylium species isolated from Allium crops

  • Chang-Gi Back;You-Kyung Han;Walftor Bin Dumin;Jong-Han Park;Ji-Won Han;Yeoung-Seok Bae
    • Korean Journal of Agricultural Science
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    • v.49 no.1
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    • pp.113-120
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    • 2022
  • Leaf blight disease caused by Stemphylium species is an important disease in Allium crops, specifically onion, garlic and welsh onion. In 2018, leaf blight symptoms were severe and damaged onion and garlic in Jeonnam province in Korea. In addition, small purple spots on garlic burbs were observed in a post-harvest storage warehouse. Several Stemphylium isolates were isolated from diseased leaves from the field and from garlic bulb samples and were analyzed in terms of homology and the phylogenetic relationship based on the internal transcribed spacer region and calmodulin gene sequence. The results showed that among three Stemphylium species identified, S. vesicarium is most prevalent on onion and garlic. S. eturmiunum was for the first time identified as pathogenic to onion and garlic, whereas S. solani was found in welsh onion crops. Although these isolates grew well at the optimum temperature at 20 - 25℃, they could also grow at low temperatures of 10 - 15℃. A pathogenicity test was conducted using S. vesicarium and S. eturmiunum on onion and garlic respectively. These results showed that two Stemphylium species were highly virulent with cross pathogenicity in onion and garlic. The results of this study can support the biological characterization of Stemphylium species in Korea. Moreover, further research will need to develop fungicide application strategies for onion and garlic crops.

Phylogenetic placement of thermophilic ammonium-tolerant bacteria and their distribution in various composts

  • Kazutaka Kuroda
    • Animal Bioscience
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    • v.36 no.4
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    • pp.671-678
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    • 2023
  • Objective: Previous studies isolated the thermophilic ammonium-tolerant (TAT) bacterium Bacillus sp. TAT105 that grew in composting swine manure with the assimilation of ammonium nitrogen and reduced ammonia emissions during composting. Those studies also investigated the potential for applications of TAT105 to composting. It was observed that the concentration of TAT bacteria, phylogenetically close to TAT105, increased during composting. The objectives of this study were to identify the phylogenetic placement of these TAT bacteria and investigate their distribution in various composts. Methods: The phylogenetic placement of TAT105 was examined based on the sequence of 16S ribosomal RNA gene. The genomic DNA homology between TAT105 and the type strains of bacterial species that were phylogenetically close to TAT105 were examined by DNA-DNA hybridization. Moreover, the tolerances of these strains to NH4Cl and NaCl were analyzed using a cultivation method. Concentrations of TAT bacteria in various composts were evaluated using an agar medium specific to TAT bacteria and polymerase chain reaction followed by restriction fragment length polymorphism analysis. Results: TAT105 was most closely related to Bacillus thermolactis and Bacillus kokeshiiformis. Many variants of these species have been detected in various environments, including composts. The type strains of these species displayed TAT characteristics that were similar to those of TAT105. Among the composts examined in this study, TAT bacteria were detected at high concentrations (105 to 109 colony forming units per gram of dry matter) in most of the composts made from cattle manure, swine manure, bark, and excess sludge. Conclusion: TAT bacteria comprised B. thermolactis, B. kokeshiiformis, and their phylogenetically close relatives. They were considered to be adaptable to composting of some certain materials, and a favorable target for searching for strains with some useful function that could be applied to composting of these materials.

Inhibition of Melanosome Transport by Inducing Exon Skipping in Melanophilin

  • Jin Young Kim;Seon-Young Han;Kiho Sung;Jeong Yeon Seo;Cheol Hwan Myung;Chan Song Jo;Jee Hoe Yoon;Ji Yun Park;Jae Sung Hwang
    • Biomolecules & Therapeutics
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    • v.31 no.4
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    • pp.466-472
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    • 2023
  • Exon skipping is an efficient technique to inhibit specific gene expression induced by a short-sequence peptide nucleic acid (PNA). To date, there has been no study on the effects of PNA on skin pigmentation. In melanocytes, the tripartite complex is responsible for the transport of mature melanosomes from the nucleus to the dendrites. The tripartite complex is composed of Rab27a, Mlph (Melanophilin), and Myosin Va. Defects in the protein Mlph, a melanosome transport-related protein, are known to cause hypopigmentation. Our study shows that Olipass peptide nucleic acid (OPNA), a cell membrane-permeable PNA, targets exon skipping in the Mlph SHD domain, which is involved in Rab27a binding. Our findings demonstrate that OPNA induced exon skipping in melan-a cells, resulting in shortened Mlph mRNA, reduced Mlph protein levels, and melanosome aggregation, as observed by microscopy. Therefore, OPNA inhibits the expression of Mlph by inducing exon skipping within the gene. These results suggest that OPNA, which targets Mlph, may be a potential new whitening agent to inhibit melanosome movement.

Genetic Characteristics of Extended-Spectrum Beta-Lactamase-Producing Salmonella Isolated from Retail Meats in South Korea

  • Haiseong Kang;Hansol Kim;Hyochin Kim;Ji Hye Jeon;Seokhwan Kim;Yongchjun Park;Soon Han Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.5
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    • pp.1101-1108
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    • 2024
  • Earlier studies have validated the isolation of extended-spectrum beta-lactamase-producing Salmonella (ESBL-Sal) strains from food. While poultry is recognized as a reservoir for Salmonella contamination, pertinent data regarding ESBL-Sal remains limited. Consequently, the Ministry of Food and Drug Safety has isolated Salmonella spp. from retail meat and evaluated their antibiotic susceptibility and genetic characteristics via whole-genome sequencing. To further elucidate these aspects, this study investigates the prevalence, antibiotic resistance profiles, genomic characteristics, and homology of ESBL-Sal spp. obtained from livestock-derived products in South Korean retail outlets. A total of 653 Salmonella spp. were isolated from 1,876 meat samples, including 509 beef, 503 pork, 555 chicken, and 309 duck samples. The prevalence rates of Salmonella were 0.0%, 1.4%, 17.5%, and 28.2% in the beef, pork, chicken, and duck samples, respectively. ESBL-Sal was exclusively identified in poultry meat, with a prevalence of 1.4% in the chicken samples (8/555) and 0.3% in the duck samples (1/309). All ESBL-Sal strains carried the blaCTX-M-1 gene and exhibited resistance to ampicillin, ceftiofur, ceftazidime, nalidixic acid, and tetracycline. Eight ESBL-Sal isolates were identified as S. Enteritidis with sequence type (ST) 11. The major plasmid replicons of the Enteritidis-ST11 strains were IncFIB(S) and IncFII(S), carrying antimicrobial resistance genes (β-lactam, tetracycline, and aminoglycoside) and 166 virulence factor genes. The results of this study provide valuable insights for the surveillance and monitoring of ESBL-Sal in South Korean food chain.

Molecular characterization of gonadotropin-releasing hormone (GnRH) genes and their role in reproductive system of Pangasius species

  • Amirah Syafiqah Zamri;Fatin Nabilah Sahadan;Zarirah Zulperi;Fadhil Syukri;Yuzine Esa
    • Fisheries and Aquatic Sciences
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    • v.27 no.6
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    • pp.366-378
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    • 2024
  • Application of commercial hormone failed to promote breeding in certain Pangasius species due to the differences of gonadotropin-releasing hormone specific peptide with species-specific bioactivities. Gonadotropin-releasing hormone (GnRH) is a hypothalamic decapeptide in the reproductive system that plays a crucial role in the regulation of reproductive processes. This study was performed to determine and analyse the GnRH genes from commercially important Pangasius sp., Pangasianodon hypophthalmus and Pangasius nasutus. The GnRH1 and GnRH2 genes were amplified and cloned into TOPO vector, followed by phylogenetic analysis of a complete open reading frame (ORF) of GnRH genes. The GnRH1 and GnRH2 genes of P. hypophthalmus and P. nasutus were detected at 300 bp and 360 bp, encoded for 81 and 87 amino acids, respectively. Amino acid sequence identities revealed high homology of P. hypophthalmus and P. nasutus GnRH1 and GnRH2 genes in comparison with other fish and vertebrates. Phylogenetic tree showed that fish from various families were aggregated into a group of the same order due to their highest identity similarities. It revealed that the vertebrate formed clusters and are grouped according to their GnRH decapeptide and GnRH-associated peptide (GAP) region, indicating a close relationship among GnRH decapeptide and GAP in different vertebrate species.

Genes of Wild Rice (Oryza grandiglumis) Induced by Wounding and Yeast Extract (상처와 효모추출물 처리조건에서 유발되는 야생벼 유전자 스크린)

  • Shin, Sang-Hyun;Im, Hyun-Hee;Lee, Jai-Heon;Kim, Doh-Hoon;Chung, Won-Bok;Kang, Kyung-Ho;Cho, Sung-Ki;Shin, Jeong-Sheop;Chung, Young-Soo
    • Journal of Life Science
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    • v.14 no.4
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    • pp.650-656
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    • 2004
  • Oryza grandiglumis (CCDD, 2n=48), one of the wild rice species, has been known to possess fungal-,bacterial-, and insect-resistance against sheath blight, rice blast, bacterial leaf blight and brown plant hopper (Nilaparvata lugens). To rapidly isolate differentially expressed genes responding to fungal and wounding stress, wounding and yeast extract were treated to O. grandiglumis for 24 hrs. Suppression subtractive hybridization (SSH) method was used to obtain differentially expressed genes from yeast extract and wounding treated plants. Seven hundreds and seventy six clones were obtained by subcloning PCR product, and colony array and screening were carried out using radio-isotope labeled cDNA probes prepared from the wounding and yeast extract treated plants. One hundred and fifteen colonies were confirmed as true positive ones. Average insert size of the clones were ranged from 400 bp to 700 bp and all the inserts were sequenced. To decide the identity of those clones, sequences were analyzed by sequence homology via GenBank database. The homology search result showed that 68 clones were matched to the genes with known function; 16 were related to primary metabolism, 5 to plant retrotransposons, 5 to defense related metallothionein-like genes. In addition to that, others were matched to various genes with known function in amino acid synthesis and processing, membrane transport, and signal transduction, so on. In northern blot analysis, induced expressions of ogwfi-161, ogwfi-646, ogwfi-663, and ogwfi-695 by wounding and yeast extract treatments were confirmed. The result indicates that SSH method is very efficient for rapid screening of differentially expressed genes.

Molecular Characterization and Phylogenetic Analysis of Season Influenza Virus Isolated in Busan during the 2006-2008 Seasons (부산지역에서 유행한 계절인플루엔자바이러스의 유전자 특성 및 계통분석('06-'08 절기))

  • Park, Yon-Koung;Kim, Nam-Ho;Choi, Seung-Hwa;Lee, Mi-Oak;Min, Sang-Kee;Kim, Seong-Joon;Cho, Kyung-Soon;Na, Young-Nan
    • Journal of Life Science
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    • v.20 no.3
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    • pp.365-373
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    • 2010
  • To monitor newly emerged influenza virus variants and to investigate the prevalence pattern, our laboratory performed isolation of the viruses from surveillance sentinel hospitals. In the present study, we analysed influenza A/H1N1, A/H3N2, B viruses isolated in Busan during the 2006/07 and 2007/08 seasons by sequence analysis of the hemagglutinin (HA1 subunit) and neuraminidase (NA) genes. The isolates studied here were selected by the stratified random sample method from a total of 277 isolates, in which 15 were A/H1N1, 16 were A/H3N2 and 29 were B. Based on the phylogenetic tree, the HA1 gene showed that A/H1N1 isolates had a 96.7% to 97.7% homology with the A/Brisbane/59/2007, A/H3N2 isolates had a 98.4% to 99.7% homology with the A/Brisbane/10/2007, and B isolates had a 96.5% to 99.7% homology with the B/Florida/4/2006(Yamagata lineage), which are all the vaccine strains for the Northern Hemisphere in 2008~2009 season. In the case of the NA gene, A/H1N1 isolates had 97.8% to 98.5% homologies, A/H3N2 isolates had 98.9% to 99.4% homologies, and B isolates had 98.9% to 100% homologies with each vaccine strain in the 2008~2009 season, respectively. Characterization of the hemagglutinin gene revealed that amino acids at the receptor-binding site and N-linked glycosylation site were highly conserved. These results provide useful information for the control of influenza viruses in Busan and for a better understanding of vaccine strain selection.

Characterization of the DGAT1 Gene in the Korean Holstein Dairy Cattle Population (한국 Holstein종 유우집단의 DGAT1 유전자의 특성분석)

  • Son, Ji-Young;Jeong, Hang-Jin;Yu, Seong-Lan;Lee, Jun-Heon;Do, Chang-Hee;Ryoo, Seung-Heui;Sang, Byung-Chan
    • Korean Journal of Agricultural Science
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    • v.36 no.2
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    • pp.167-177
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    • 2009
  • This study was conducted to characterize the DGAT1 gene in Korean Holstein dairy cattle population and examine the relationship of DGAT1 polymorphisms with milk yield and milk fat yield for the genetic improvement of Korean Holstein dairy cattle. Results indicated that the 411 bp PCR products were successfully amplified by DGAT1 specific primers. Sequence analysis indicated that the DGTA1 Q allele had AUG (Lysine, K) nucleotide sequences in 216-218 bp and q allele had GCG (Alanine, A) sequences in the same position. Nucleotide sequence homology between the DGAT1 sequences generated in this study showed 100% homology with bovine DGAT1 sequences in the NCBI database. The genotype frequencies of DGAT1 QQ, Qq, and qq were 16.43, 36.43, and 47.14%, respectively, in Korean Holstein dairy cattle population. The observed Q and q allele frequencies were 0.35 and 0.65, respectively. Statistically significant (P<0.05) results were identified for milk yield and milk fat yield for the DGAT1 genotypes. The Qq genotype Holsteins have significantly higher milk yield and milk fat yield than those of the QQ and qq genotype Holsteins(P<0.05).

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Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea (우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석)

  • ;Richard M. Bostock
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.77-96
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    • 1995
  • To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

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