• Title/Summary/Keyword: sequence homology

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Characterization and functional analysis of a myo-inositol 1-phosphate synthase cDNA in sesame (Sesamum indicum L.) (참깨 myo-inositol 1-phosphate synthase 유전자의 특성과 기능분석에 관한 연구)

  • 진언호;천재안;정정한
    • Journal of Life Science
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    • v.13 no.4
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    • pp.383-389
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    • 2003
  • A cDNA (SeMIPS) encoding myo-inositol 1-phosphate synthase has been isolated from developing sesame (Sesamum indicum L. cv. Dan-Baek) seeds and its structure and function analyzed. The SeMIPS protein was highly homologous with those from plant species (88-94%), while a much lower degree of sequence homology (60%) was found with that of human. The functional domains commonly found in MIPS protein were identified and their amino acid residues were compared with each other. Northern blot indicated that the expression of the SeMIPS gene might be organ-specifically regulated. A complementation assay based on a yeast mutant system confirmed that the SeMIPS gene encodes a myo-inositol 1-phosphate synthase (MIPS) of sesame by showing functional expression of the SeMIPS cDNA in the yeast mutants containing the disrupted INO1 gene.

Downregulation of PyHRG1, encoding a novel secretory protein in the red alga Pyropia yezoensis, enhances heat tolerance

  • Han, Narae;Wi, Jiwoong;Im, Sungoh;Lim, Ka-Min;Lee, Hun-Dong;Jeong, Won-Joong;Kim, Geun-Joong;Kim, Chan Song;Park, Eun-Jeong;Hwang, Mi Sook;Choi, Dong-Woog
    • ALGAE
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    • v.36 no.3
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    • pp.207-217
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    • 2021
  • An increase in seawater temperature owing to global warming is expected to substantially limit the growth of marine algae, including Pyropia yezoensis, a commercially valuable red alga. To improve our knowledge of the genes involved in the acquisition of heat tolerance in P. yezoensis, transcriptomes sequences were obtained from both the wild-type SG104 P. yezoensis and heat-tolerant mutant Gy500. We selected 1,251 differentially expressed genes that were up- or downregulated in response to the heat stress condition and in the heat-tolerant mutant Gy500, based on fragment per million reads expression values. Among them, PyHRG1 was downregulated under heat stress in SG104 and expressed at a low level in Gy500. PyHRG1 encodes a secretory protein of 26.5 kDa. PyHRG1 shows no significant sequence homology with any known genes deposited in public databases to date. However, PyHRG1 homologs were found in other red algae, including other Pyropia species. When PyHRG1 was introduced into the single-cell green alga Chlamydomonas reinhardtii, transformed cells overexpressing PyHRG1 showed severely retarded growth. These results demonstrate that PyHRG1 encodes a novel red algae-specific protein and plays a role in heat tolerance in algae. The transcriptome sequences obtained in this study, which include PyHRG1, will facilitate future studies to understand the molecular mechanisms involved in heat tolerance in red algae.

Expression and Purification of Transmembrane Protein MerE from Mercury-Resistant Bacillus cereus

  • Amin, Aatif;Sarwar, Arslan;Saleem, Mushtaq A.;Latif, Zakia;Opella, Stanley J.
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.274-282
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    • 2019
  • Mercury-resistant ($Hg^R$) bacteria were isolated from heavy metal polluted wastewater and soil collected near to tanneries of district Kasur, Pakistan. Bacterial isolates AZ-1, AZ-2 and AZ-3 showed resistance up to $40{\mu}g/ml$ against mercuric chloride ($HgCl_2$). 16S rDNA ribotyping and phylogenetic analysis were performed for the characterization of selected isolates as Bacillus sp. AZ-1 (KT270477), Bacillus cereus AZ-2 (KT270478) and Bacillus cereus AZ-3 (KT270479). Phylogenetic relationship on the basis of merA nucleotide sequence confirmed 51-100% homology with the corresponding region of the merA gene of already reported mercury-resistant Gram-positive bacteria. The merE gene involved in the transportation of elemental mercury ($Hg^0$) via cell membrane was cloned for the first time into pHLV vector and transformed in overexpressed C43(DE3) E. coli cells. The recombinant plasmid (pHLMerE) was expressed and the native MerE protein was obtained after thrombin cleavage by size exclusion chromatography (SEC). The purification of fusion/recombinant and native protein MerE by Ni-NTA column, dialysis and fast protein liquid chromatography (FPLC/SEC) involved unfolding/refolding techniques. A small-scale reservoir of wastewater containing $30{\mu}g/ml$ of $HgCl_2$ was designed to check the detoxification ability of selected strains. It resulted in 83% detoxification of mercury by B. cereus AZ-2 and B. cereus AZ-3, and 76% detoxification by Bacillus sp. AZ-1 respectively (p < 0.05).

The first case report on Vibrio atlanticus infection in cultured tiger puffer, Takifugu rubripes (우리나라 양식 자주복, Takifugu rubripes (Temminck & Schlegel, 1850)에서 발생한 Vibrio atlanticus의 첫 감염 사례 보고)

  • Lee, Nam-Sil;Cho, Miyoung;Jung, Sung-hee;Won, Kyoungmi
    • Journal of fish pathology
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    • v.31 no.2
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    • pp.87-92
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    • 2018
  • A case of disease was occurred in tiger puffer, Takifugu rubripes, reared in circulated system (water temperature was about $20^{\circ}C$) on land in Korea. Diseased fish showed a unique external sign, the cloudy skin ulcer in various sizes. We investigated the cause of disease, and isolated one pure cultured bacterium, TP01. This had 99.7% homology with Vibrio atlanticus by sequence analysis of the 16S rRNA and rpoA genes. TP01 showed some differences with other reported V. atlanticus strains in the biochemical characteristics as like Voges-Proskauer test and gelatin hydrolysis.

Blood transcriptome resources of chinstrap (Pygoscelis antarcticus) and gentoo (Pygoscelis papua) penguins from the South Shetland Islands, Antarctica

  • Kim, Bo-Mi;Jeong, Jihye;Jo, Euna;Ahn, Do-Hwan;Kim, Jeong-Hoon;Rhee, Jae-Sung;Park, Hyun
    • Genomics & Informatics
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    • v.17 no.1
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    • pp.5.1-5.9
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    • 2019
  • The chinstrap (Pygoscelis antarcticus) and gentoo (P. papua) penguins are distributed throughout Antarctica and the sub-Antarctic islands. In this study, high-quality de novo assemblies of blood transcriptomes from these penguins were generated using the Illumina MiSeq platform. A total of 22.2 and 21.8 raw reads were obtained from chinstrap and gentoo penguins, respectively. These reads were assembled using the Oases assembly platform and resulted in 26,036 and 21,854 contigs with N50 values of 929 and 933 base pairs, respectively. Functional gene annotations through pathway analyses of the Gene Ontology, EuKaryotic Orthologous Groups, and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were performed for each blood transcriptome, resulting in a similar compositional order between the two transcriptomes. Ortholog comparisons with previously published transcriptomes from the $Ad{\acute{e}}lie$ (P. adeliae) and emperor (Aptenodytes forsteri) penguins revealed that a high proportion of the four penguins' transcriptomes had significant sequence homology. Because blood and tissues of penguins have been used to monitor pollution in Antarctica, immune parameters in blood could be important indicators for understanding the health status of penguins and other Antarctic animals. In the blood transcriptomes, KEGG analyses detected many essential genes involved in the major innate immunity pathways, which are key metabolic pathways for maintaining homeostasis against exogenous infections or toxins. Blood transcriptome studies such as this may be useful for checking the immune and health status of penguins without sacrifice.

A Role of YlBud8 in the Regulation of Cell Separation in the Yeast Yarrowia lipolytica

  • Li, Yun-Qing;Xue, Qing-Jie;Yang, Yuan-Yuan;Wang, Hui;Li, Xiu-Zhen
    • Journal of Microbiology and Biotechnology
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    • v.29 no.1
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    • pp.141-150
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    • 2019
  • The spatial landmark protein Bud8 plays a crucial role in bipolar budding in the budding yeast Saccharomyces cerevisiae. The unconventional yeast Yarrowia lipolytica can also bud in a bipolar pattern, but is evolutionarily distant from S. cerevisiae. It encodes the protein YALI0F12738p, which shares the highest amino acid sequence homology with S. cerevisiae Bud8, sharing a conserved transmembrane domain at the C-terminus. Therefore, we named it YlBud8. Deletion of YlBud8 in Y. lipolytica causes cellular separation defects, resulting in budded cells remaining linked with one another as cell chains or multiple buds from a single cell, which suggests that YlBud8 may play an important role in cell separation, which is distinct from the function of Bud8 in S. cerevisiae. We also show that the YlBud8-GFP fusion protein is located at the cell membrane and enriched in the bud cortex, which would be consistent with a role in the regulation of cell separation. The coiled-coil domain at the N-terminus of YlBud8 is important to the correct localization and function of YlBud8, as truncated proteins that do not contain the coiled-coil domain cannot rescue the defects observed in $Ylbud8{\Delta}$. This finding suggests that a new signaling pathway controlled by YlBud8 via regulation of cell separation may exist in Y. lipolytica.

Prevalence and pathologic study of porcine epidemic diarrhea in Jeju (제주도의 돼지유행성설사 발생 상황과 병리학적 고찰)

  • Kim, Hyeon-Ju;Yang, Hyoung-Seok;Kang, Sang Chul;Kim, Jae-Hoon
    • Korean Journal of Veterinary Research
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    • v.61 no.4
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    • pp.30.1-30.11
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    • 2021
  • From April 2014 to September 2015, 153 piglets from 52 farms in Jeju were diagnosed with porcine epidemic diarrhea (PED). The major PED cases were focused on suckling piglets (144 piglets, 94.1%), particularly in 1-7-day-old piglets. Histopathologically, severe villous atrophy was observed in the small intestine, especially in the jejunum and ileum. The mean villous height to crypt depth ratios of the jejunum and ileum were 1.4:1 and 1.5:1, respectively. The major histopathologic findings of the small intestine were cytoplasmic vacuolation, cuboidalization, squamation, and exfoliation of the mucosal enterocytes in the villi. The cytoplasmic vacuolations in the enterocytes were the most prevalent lesions in the small intestine and were more severe in the ileum than in the jejunum. According to immunohistochemistry methods, the PED virus (PEDV) antigens were presented in the cytoplasms of the enterocytes, and were distributed more prevalently in the ileum than in the jejunum. PEDV antigens were also detected in the colon of 26 piglets (19.5%). Sequence comparison and phylogenetic analysis indicated that 12 PEDV had more than a 98.9% homology with each other. These PEDV strains were highly homologous with the genogroup 2 North American group.

Molecular characterization and functional annotation of a hypothetical protein (SCO0618) of Streptomyces coelicolor A3(2)

  • Ferdous, Nadim;Reza, Mahjerin Nasrin;Emon, Md. Tabassum Hossain;Islam, Md. Shariful;Mohiuddin, A.K.M.;Hossain, Mohammad Uzzal
    • Genomics & Informatics
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    • v.18 no.3
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    • pp.28.1-28.9
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    • 2020
  • Streptomyces coelicolor is a gram-positive soil bacterium which is well known for the production of several antibiotics used in various biotechnological applications. But numerous proteins from its genome are considered hypothetical proteins. Therefore, the present study aimed to reveal the functions of a hypothetical protein from the genome of S. coelicolor. Several bioinformatics tools were employed to predict the structure and function of this protein. Sequence similarity was searched through the available bioinformatics databases to find out the homologous protein. The secondary and tertiary structure were predicted and further validated with quality assessment tools. Furthermore, the active site and the interacting proteins were also explored with the utilization of CASTp and STRING server. The hypothetical protein showed the important biological activity having with two functional domain including POD-like_MBL-fold and rhodanese homology domain. The functional annotation exposed that the selected hypothetical protein could show the hydrolase activity. Furthermore, protein-protein interactions of selected hypothetical protein revealed several functional partners those have the significant role for the bacterial survival. At last, the current study depicts that the annotated hypothetical protein is linked with hydrolase activity which might be of great interest to the further research in bacterial genetics.

Allergenicity and toxicity evaluation of the PAT protein expressed in herbicide-tolerant genetically modified Zoysia japonica (제초제저항성 GM 잔디에서 발현된 PAT 단백질의 알레르겐 유발 가능성 및 독성 평가)

  • Jeong, Hye-Rin;Sun, Hyeon-Jin;Kang, Ji-Nam;Kang, Hong-Gyu;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.47 no.4
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    • pp.316-323
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    • 2020
  • This study aimed to evaluate the potential allergenicity and oral toxicity of the phosphinothricin acetyltransferase (PAT) protein expressed in Zoysia japonica, a herbicide-tolerant genetically modified (GM) zoysiagrass. In silico analysis of PAT showed no similarities with any known allergenic or toxic proteins, with <35% amino acid sequence homology with known allergens across a length of 80 amino acids and no continuous eight amino acid identity with known allergens. The PAT protein expressed in Z. japonica degraded very rapidly in the simulated gastric fluid in the presence of pepsin, and, no glycosylation of PAT was observed. The oral toxicity test revealed no mortality or toxic effect in mice following PAT administration at 4,000 mg/kg body weight. Our findings indicate that the PAT protein expressed in Zoysia japonica does not exhibit allergenic or toxic properties.

Screening and Purification of a Novel Antibacterial Peptide, cgCAFLP, Against Skin Pathogens from the Extract of the Pacific Oyster Crassostrea gigas from Buan in Korea (부안산 참굴(Crassostrea gigas) 추출물로부터 피부 상재균에 대한 새로운 항균 펩타이드, cgCAFLP의 탐색 및 정제)

  • Lee, Ji-Eun;Seo, Jung-Kil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.6
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    • pp.927-937
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    • 2021
  • This study was performed to screen the antimicrobial activities of the extract from the Pacific oyster Crassostrea gigas against skin pathogens and to purify the relevant antibacterial peptide. The acidified extract showed potent antibacterial activities against gram-positive and gram-negative bacteria but showed no activity against Candida albicans and no significant cell toxicity. Among acne-causing pathogens, the acidified extract showed potent antibacterial activity only against Staphylococcus aureus, and its antibacterial activity was completely abolished by treatment with trypsin or chymotrypsin, and was inhibited by salt treatment. The acidified extract showed strong DNA-binding ability but did not show bacterial membrane permeabilizing ability. Based on antimicrobial activity screening and cytotoxic effects, a novel antibacterial peptide was purified from the acidified gill extract using solid-phase extraction, cation-exchange, and reversed-phase HPLC. The resulting peptide had a molecular weight of 4800.8 Da and showed partial sequence homology with the carbonic anhydrase 4 (CA4) protein in the hard-shelled mussel. Overall, we purified a novel antibacterial peptide, named cgCAFLP, which is related to carbonic anhydrase 4 (CA4) protein, against skin pathogens. Our results suggest that the Pacific oyster extract could be used as an additive to control some acne-related skin pathogens (S. aureus).