• Title/Summary/Keyword: tRNA gene

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A report of 5 unrecorded bacterial species of the Deinococcus genus in Korea

  • Lee, Jae-Jin;Kang, Myung-Suk;Joo, Eun Sun;Kim, Myung Kyum
    • Journal of Species Research
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    • v.5 no.1
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    • pp.22-26
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    • 2016
  • Five bacterial strains designated DY37, BS333, JJ521, BM1, and DG13-2 were assigned to the genus Deinococcus were isolated from forest soil samples collected from Deogyusan, Busan, Changwon, and Seoul of South Korea. The isolates were Gram-staining negative or positive, and pale pink- or red-pigmented, short-rod shaped. Phylogenetic analysis based on 16S rRNA gene sequence revealed that strains DY37, BS333, JJ521, BM1, and DG13-2 were most closely related to Deinococcus aquatilis CCM $7524^T$ (with 99.0% similarity), D. ficus CC-FR2-$10^T$ (100.0%), D. grandis KS $0485^T$ (99.2%), D. roseus TDMA-$uv51^T$ (98.9%), and D. yunweiensis $YIM007^T$(100.0%), respectively. These 5 species have never been proposed in Korea; therefore 5 species of 1 genera in the family Deinococcaceae in the order Deinococcales within the class Deinococci are reported for proteobacterial species found in Korea.

Cohnella damensis sp. nov., a Motile Xylanolytic Bacteria Isolated from a Low Altitude Area in Tibet

  • Luo, Xuesong;Wang, Zhang;Dai, Jun;Zhang, Lei;Fang, Chengxiang
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.410-414
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    • 2010
  • A bacterial strain, 13-$25^T$ with xylanolytic activity isolated from a single soil sample, was characterized with respect to its phenetic and phylogenetic characteristics. The cells of the isolate are Gram-staining variable rods, but spore formation was not observed. This strain is catalase- and oxidase-positive, and able to degrade starch and xylan. The predominant fatty acids are anteiso-$C_{15:0}$, $C_{16:0}$, and iso-$C_{16:0}$. The major respiratory quinone is menaquinone 7(MK-7), with a polar lipid profile consistent with the genus Cohnella. The DNA G+C content is 54.3 mol%. The 168 rRNA gene sequence analysis indicates that this organism belongs to the genus Cohnella, with Cohnella panacarvi as the closest phylogenetic neighbor. Low levels of 168 rRNA gene sequence similarity (<97.0%) with respect to other taxa with published names and the identification of distinctive phenetic features in the isolate indicate that the strain 13-$25^T$ represents a novel species of the genus Cohnella, for which the name Cohnella damensis sp. novo is proposed. The type strain is 13-$25^T$ (=CCTCC AB $208103^T$=KCTC $13422^T$).

Pedobacter xinjiangensis sp. nov., from the Desert, Xinjiang

  • Tang, Yali;Wang, Yang;Ji, Shanming;Zhang, Kundi;Dai, Jun;Zhang, Lei;Peng, Fang;Fang, Chengxiang
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.397-402
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    • 2010
  • A Gram-negative, rod-shaped, gliding, aerobic bacterium, designated $12157^T$, was isolated from the desert of Xinjiang, China and subjected to a polyphasic taxonomic study. The strain $12157^T$ grew optimally at pH 7.0 and $30^{\circ}C$. MK-7 was the predominant respiratory menaquinone. The DNA G+C content was 42.0 mol%. Phylogenetic analysis based on the 16S rRNA gene sequences showed that the isolate was mostly related to members of the genus Pedobacter, with similarities ranging from 90.0% to 93.7%. Phylogenetic evidence and the results of phenotypic and genotypic analyses support the establishment of a novel species, Pedobacter xinjiangensis sp. nov., with strain $12157^T$ (=CCTCC AB $208092^T$=NRRL B-$51338^T$) as the type strain.

Editing of Genomic TNFSF9 by CRISPR-Cas9 Can Be Followed by Re-Editing of Its Transcript

  • Lee, Hyeon-Woo
    • Molecules and Cells
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    • v.41 no.10
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    • pp.917-922
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    • 2018
  • The CRISPR-Cas system is a well-established RNA-guided DNA editing technique widely used to modify genomic DNA sequences. I used the CRISPR-Cas9 system to change the second and third nucleotides of the triplet $T{\underline{CT}}$ of human TNSFSF9 in HepG2 cells to $T{\underline{AG}}$ to create an amber stop codon. The $T{\underline{CT}}$ triplet is the codon for Ser at the $172^{nd}$ position of TNSFSF9. The two substituted nucleotides, AG, were confirmed by DNA sequencing of the PCR product followed by PCR amplification of the genomic TNFSF9 gene. Interestingly, sequencing of the cDNA of transcripts of the edited TNFSF9 gene revealed that the $T{\underline{AG}}$ had been re-edited to the wild type triplet $T{\underline{CT}}$, and 1 or 2 bases just before the triplet had been deleted. These observations indicate that CRISPR-Cas9-mediated editing of bases in target genomic DNA can be followed by spontaneous re-editing (correcting) of the bases during transcription.

Effects of 2-deoxy-D-glucose and quercetin on the expression of osteonectin and osteopontin during the differentiation of irradiated MC3T3-El osteoblastic cells

  • Yu, Su-Kyoung;Koh, Kwang-Joon;Kim, Kyoung-A
    • Imaging Science in Dentistry
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    • v.38 no.4
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    • pp.195-202
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    • 2008
  • Purpose : To characterize the effects of 2-deoxy-D-glucose (2-DG) and quercetin (QCT) on gene expression of osteonectin (ON) and osteopontin (OP) in irradiated MC3T3-El cells. Materials and Methods : When MC3T3-El osteoblastic cells had reached 70-80% confluence, cultures were transferred to a differentiating medium supplemented with 5 mM 2-DG or 10 ${\mu}M$ QCT and then irradiated with 2, 4, 6, and 8 Gy. At various times after irradiation, the cells were analyzed for the expression of bone mineralization genes such as ON and OP. Results : The mRNA expression of both ON and OP was increased according to the culture time in the differentiation medium, and the increase of the genes peaked at 14 days after the differentiation induction. In the case of OP, the increase of mRNA expression was maintained to 28 days after the differentiation, while the mRNA level of ON was reduced to the basal level at the same time. Irradiation adding 2-DG showed a significant peak value in the expression pattern of ON at 4 Gy 7 days after irradiation. Irradiation adding QCT increased the mRNA expression of ON and OP in a dose-dependant manner, but irradiation adding 2-DG did not show any differences between the control and experiments 14 days after irradiation. Irradiation adding QCT increased significantly the expression patterns of ON 21 days after irradiation. Conclusion : The results showed that QCT acted as a radiosensitizer in the gene expression of ON and OP during differentiation of the late stage of irradiated MC3T3-E1 osteoblastic cells in vitro. (Korean J Oral Maxillofac Radiol 2008; 38: 195-202)

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Complete genome sequence of biofilm-producing strain Staphylococcus xylosus S170 (생물막 생성 Staphylococcus xylosus S170 균주의 유전체 분석연구)

  • Hong, Jisoo;Roh, Eunjung
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.167-168
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    • 2018
  • Here we report the complete genome sequence of Staphylococcus xylosus S170, strong biofilm-producing strain, which comprised a single circular 2,910,005 bp chromosome and 32.97% G + C content. The genome included 2,674 protein-coding sequences, 22 rRNA genes, and 57 tRNA genes. Gene analysis of S. xylosus S170 could contribute to better understanding of biofilm-forming mechanisms.

The innate immune response transcription factor Bombyx mori Relish1 induces high-level antimicrobial peptides in silkworm

  • Kim, Seong-Wan;Kim, Seong-Ryul;Goo, Tae-Won;Choi, Kwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.37 no.2
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    • pp.49-54
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    • 2018
  • To artificially enhance antimicrobial peptide expression in Bombyx mori, we constructed genetically engineered silkworms overexpressing Rel family transcription factor. The truncated BmRelish1 (BmRelish1t) gene contained a Rel homolog domain (RHD), nuclear localization signal (NLS), acidic and hydrophobic amino acid (AHAA)-rich region, and death domain (DD), but no ankyrin-repeat (ANK) domain. The BmRelish1t gene was controlled by B. mori cytoplasmic actin 3 promoter in the PiggyBac transposon vector. Chromosome analysis of G1 generations of a transgenic silkworm with EGFP expression confirmed stable insertion of BmRelish1t. BmRelish1t gene overexpression in transgenic silkworms resulted in higher mRNA expression levels of B. mori antimicrobial peptides such as lebocin(~20.5-fold), moricin(~8.7-fold), and nuecin(~17.4-fold) than those in normal silkworms.

Biochemical and Molecular Biological Studies on the DNA Replication of Bacteriophage T7 (Bacteriophage T7의 유전자 복제기작에 관한 생화학적, 분자생물학적 특성 연구)

  • KIM Young Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.2
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    • pp.209-218
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    • 1995
  • Bacteriophage T7 gene 2.5 protein, a single-stranded DNA binding protein, has been implicated in T7 DNA replication, recombination, and repair. Purified gene 2.5 protein has been shown to interact with the phage encoded gene 5 protein (DNA polymerase) and gene 4 proteins (helicase and primase) and stimulates their activities. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth. T7 phage that contain null mutants of gene 2.5 were constructed by homologous recombination. These mutant phage $(T7\Delta2.5)$ cannot grow in Escherichia coli. After infection of E. coli with $T7\Delta2.5$, host DNA synthesis is shut off, and $T7\Delta2.5$ DNA synthesis is reduced to less than $1\%$ of wild-type phage DNA synthesis (Kim and Richardson, 1993, Proc. Natl. Aca. Sci. USA, 90, 10173-10177). A truncated gene 2.5 protein $(GP2.5-\Delta21C)$ deleted the 21 carboxyl terminal amino acids was constructed by in vitro mutagenesis. $GP2.5-\Delta21C$ cannot substitute for wild-type gene 2.5 protein in vivo; the phage are not viable and exhibit less than $1\%$ of the DNA synthesis observed in wild-type phage-infected cells. $GP2.5-\Delta21C$ has been purified to apparent homogeneity from cells overexpressing its cloned gene. Purified $GP2.5-\Delta21C$ does not physically into「act with T1 gene 4 protein as measured by affinity chromatography and immunoblot analysis. The mutant protein cannot stimulate T7 gene 4 protein activity on RNA-primed DNA synthesis and primer synthesis. These results suggest that C-terminal domain of gene 2.5 protein is essential for protein-protein interactions.

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Bacterial Diversity in a Korean Traditional Soybean Fermented Foods (Doenjang and Ganjang) by 16S rRNA Gene Sequence Analysis

  • Cho, Kye-Man;Seo, Weon-Taek
    • Food Science and Biotechnology
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    • v.16 no.2
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    • pp.320-324
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    • 2007
  • The bacterial diversity in Korean soybean-fermented foods was investigated using a PCR-based approach. 16S rRNA sequences were amplified and cloned from two different soybean-fermented foods such as doenjang (soybean paste), and ganjang (soybean sauce). Staphylococcus equorum (60.6%), Tetragenococcus halophila (21.2%), Leuconostoc mesenteroides (9.1%), Lactobacillus sakei (6.1%), and Bacillus subtilis (3.0%) were detected among clones isolated from soybean paste samples and Halanaerobium sp. (37.5%), Halanaerobium fermentans (37.5%), T. halophila (12.5%), Staphylococcus sp. (6.3%), S. equorum (3.1%), and B. subtilis (3.1%) were detected among clones isolated from soybean sauce. Our approach revealed different bacterial distributions and diversity from those previously obtained using culture-dependent methods.