• Title/Summary/Keyword: DNA homology

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Characterization of a New Leuconostoc Species Isolated from Fresh Garlic

  • Lee, Se-Hi;Choi, Jong-Hoon;Kim, Youn-Soon;Kyung, Kyu-Hang
    • Food Science and Biotechnology
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    • v.14 no.3
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    • pp.416-419
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    • 2005
  • Unknown bacterium isolated from garlic was characterized using phenotypic methods, phylogenetic analysis, DNA-DNA hybridization, and cultural methods. The strain was identified as typical leuconostoc; Gram-positive, non-sporeforming, heterofermentative, catalase-negative and spherical. Although its 16S rRNA gene sequence showed high homology to Leuconostoc argentinum DSM $8581^T$(99.8%), DNA-DNA hybridization experiments indicated it represents novel genomic species in the genus Leuconostoc. The garlic-specific leuconostoc was more resistant to antimicrobial activity of garlic compared to other common laboratory lactic acid bacteria, and was even stimulated by low concentrations (1-2%) of garlic extract supplemented in trypticase soy broth. Growth stimulation was concentration-dependent when tested with residual aqueous layer after solvent extraction of fresh whole garlic extract.

Distribution of ddr (DNA damage response) Genes among Species of Deinococcus

  • Lim, Sangyong;Jung, Sunwook;Joe, Minho;Kim, Dongho
    • Journal of Radiation Industry
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    • v.4 no.3
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    • pp.289-295
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    • 2010
  • The bacterium Deinococcus radiodurans is one of the most resistant organisms to the effects of ionizing radiation and other DNA-damaging agents. In this study, distributions of 10 ddr (DNA damage response) genes were investigated in 8 species of Deinococcus by polymerase chain reaction (PCR). We have compared the sequences of ddr genes of D. radiodurans, D. geothermalis and D. deserti, and selected primers which are suitable for the detection of ddr in different species of Deinococcus. A sequence homology search and PCR assay showed that ddrO, which encodes a global regulator of the radiation-desiccation response, was most well conserved in the Deinococcus lineage.

Evolution of CRISPR towards accurate and efficient mammal genome engineering

  • Ryu, Seuk-Min;Hur, Junseok W;Kim, Kyoungmi
    • BMB Reports
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    • v.52 no.8
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    • pp.475-481
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    • 2019
  • The evolution of genome editing technology based on CRISPR (clustered regularly interspaced short palindromic repeats) system has led to a paradigm shift in biological research. CRISPR/Cas9-guide RNA complexes enable rapid and efficient genome editing in mammalian cells. This system induces double-stranded DNA breaks (DSBs) at target sites and most DNA breakages induce mutations as small insertions or deletions (indels) by non-homologous end joining (NHEJ) repair pathway. However, for more precise correction as knock-in or replacement of DNA base pairs, using the homology-directed repair (HDR) pathway is essential. Until now, many trials have greatly enhanced knock-in or substitution efficiency by increasing HDR efficiency, or newly developed methods such as Base Editors (BEs). However, accuracy remains unsatisfactory. In this review, we summarize studies to overcome the limitations of HDR using the CRISPR system and discuss future direction.

Genetic Similarity Between Jujube Witches¡?Broom and Mulberry Dwarf Phytoplasmas Transmitted by Hishimonus sellatus Uhler

  • Cha, Byeongjin;Han, Sangsub
    • The Plant Pathology Journal
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    • v.18 no.2
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    • pp.98-101
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    • 2002
  • Using phytoplasma universal primer pair Pl and P7, a fragment of about 1.8 kb nucleotide sequences of 16S rRNA gene and 16S-23S rRNA intergenic spacer region, and a portion of 23S rRNA gene of jujube witches'broom (JWB) and mulberry dwarf(MD) phytoplasmas were determined. The nucleotide sequences of JWB and MD were 1,850 bp and 1,831 bp long, respectively. The JWB phytoplasma sequence was aligned with the homologous sequence of MD phytoplasma. Twenty-eight base insertions and nine base deletions were found in the JWB phytoplasma sequence compared with that of MD phytoplasma. The similarity of the aligned sequences of JWB and MD was 84.8%. The near-complete 16S rRNA gene DNA sequences of JWB and MD were 1,529 bp and 1,530 bp in length, respectively, and revealed 89.0% homology. The 16S-23S rRNA intergenic spacer region DNA sequences were 263 bp and 243 bp in lengths respectively, while homology was only 70% and the conserved tRNA-lle gene of JWB and MD was located into the intergenic space region between 16S-23S rRNA gene. The nucleotide sequences were 77 bp long in both JWB and MD, and showed 97.4% sequence homology. Based on the phylogenetic analysis of the two phytoplasmas, the JWB phytoplasma belongs to the Elm yellow phytoplasma group (16S rV), whereas, the MD phytoplasma belongs to the Aster yellow group (16S rI).

Detect of Hypericin (HyH) gene in Hypericum erectum in Korea and Comparison of H. perforatum in Europe (한국내 고추나물의 하이퍼리신 유전자(HyH)의 탐색과 유럽의 서양고추나물과 비교)

  • Huh, Man-Kyu
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1034-1038
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    • 2007
  • Hypericin (HyH) is a substance which is isolated a medicinal herb, Hypericum perforatum L., commonly known as St. John's Wort. Hypericum erectum is a long-lived herb that is distributed in Korea. Cloned HyH genes H. erectum of were conformed by sequencing. The cDNA Hyp-1 sequence has 732 bp with an open reading frame of 567. Thus coding for a protein of 152 amino acid residues. A BLAST re-search using the deduced nucleotide sequences in HyH gene produced significant alignments with the H. perforatum. Sequences in HyH gene showed significant homology with Rubus idaeus putative allergen Rub-i-1 mRNA, Protein sequence comparisons revealed significant homology between Hyp-1 and the phenolic oxidative coupling protein hyp-1 of H. perforatum (98%). Additionally, Hyp-1 showed sig-nificant homology with various other classes of allergens, including Pru-av-1 (62%) from Prunus avium and allergen Bet-vl-Sc3 from Betula pendula (60%). Thus, the result of this study may offer an important information to establish an assay system for chemicals of the herbal medicines for H. erectum as well as H. perforatum.

Cellular Dynamics of Rad51 and Rad54 in Response to Postreplicative Stress and DNA Damage in HeLa Cells

  • Choi, Eui-Hwan;Yoon, Seobin;Hahn, Yoonsoo;Kim, Keun P.
    • Molecules and Cells
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    • v.40 no.2
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    • pp.143-150
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    • 2017
  • Homologous recombination (HR) is necessary for maintenance of genomic integrity and prevention of various mutations in tumor suppressor genes and proto-oncogenes. Rad51 and Rad54 are key HR factors that cope with replication stress and DNA breaks in eukaryotes. Rad51 binds to single-stranded DNA (ssDNA) to form the presynaptic filament that promotes a homology search and DNA strand exchange, and Rad54 stimulates the strand-pairing function of Rad51. Here, we studied the molecular dynamics of Rad51 and Rad54 during the cell cycle of HeLa cells. These cells constitutively express Rad51 and Rad54 throughout the entire cell cycle, and the formation of foci immediately increased in response to various types of DNA damage and replication stress, except for caffeine, which suppressed the Rad51-dependent HR pathway. Depletion of Rad51 caused severe defects in response to postreplicative stress. Accordingly, HeLa cells were arrested at the G2-M transition although a small amount of Rad51 was steadily maintained in HeLa cells. Our results suggest that cell cycle progression and proliferation of HeLa cells can be tightly controlled by the abundance of HR proteins, which are essential for the rapid response to postreplicative stress and DNA damage stress.

Sero-epidemiology and genetic characterization of swine influenza virus (돼지 인플루엔자 바이러스의 혈청학적 역학조사 및 유전학적 분석)

  • Lyoo, Young-soo;Kim, Lomi
    • Korean Journal of Veterinary Research
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    • v.38 no.1
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    • pp.53-63
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    • 1998
  • Total of 1085 swine sera (1996-1997) from nation-wide were tested for the presence of antibodies to influenza A virus. Fifty nine percent of the tested sera showed seropositive by HI test. Positive sera consisted of 24--- of H3, 15--- of H1, and 20--- of the sample had both antibodies, respectively. Sera collected from various region represented 7~27--- seropositivity to H1N1, 15~25--- to H3N2, respectively. Swine influenza field isolate from nasal swab was characterized antigenically and genetically to elucidate its relatedness with other known strains of influenza A virus. The study was focused on the HA gene which is related to pathogenecity and antigenic variability of the influenza virus. By RT-PCR using influenza A/H1N1 specific primers, influenza virus H1N1 specific DNA fragment was amplified from A/Swine/Iowa/15/30(H1N1), US field isolate but not in H3N2 strain. PCR products were sequenced by dideoxy chain termination method to determine nucleotide homology with other strains of influenza A virus. The US field isolate and A/Swine/Indiana/1726/88 strain had 97--- of nucleotide homology and 98--- of amino acid homology. Based on the results obtained from this experiment, the field isolate was genetically related to A/Swine/Indiana/1726/88 and had higher homology with A/Swine/Indiana/1726/88 than with classical swine influenza virus, A/Swine/Iowa/15/30. The field isolate had no amino acid changes at the antigenic site compare to that of the A/Swine/Indiana/1726/88. The proteolytic enzyme cleavage site between HA1 and HA2 had no alteration and the amino acid arginine was intact. There is no evidence has been found that the field isolate has genetic shift or genetic drift which might altered antigenic determinant.

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Cloning and Characterization of the Paraquat Resistance-Related Genes from Ochrobactrum anthropi JW-2 (Ochrobactrum anthropi JW-2 유래의 Paraquat 내성유전자 PqrA의 주변 유전자군 분석)

  • Bae Eun-Kyung;Lee Hyo-Shin;Won Sung-Hye;Lee Byung-Hyun
    • Microbiology and Biotechnology Letters
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    • v.34 no.1
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    • pp.15-22
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    • 2006
  • A 4,971 bp chromosomal DNA fragment containing the pqrA, paraquat resistance gene, was cloned from Ochrobactrum anthropi JW-2, and the complete nucleotide sequence was determined. Nucleotide and deduced amino acid sequences of the fragment revealed the presence of 4 complete ORFs (orf2, pqrA, orf3, orf4) and two incomplete ORFs(orf1, orf5). Orf1, pqrA, orf4 and orf5 exists at the direct strand but orf2 and orf3 exists at the reverse complementary strand. Orf1 which of incomplete sequences without start codon shares homology with ATP binding region of the response regulator receiver. Orf2 shares high homology with members of the tetR family of transcriptional repressor which have a helix-turn-helix (H-T-H) motif. Therefore, the orf2 is predicted as a transcriptional repressor of pqrA and is designated as pqrR2. Orf3 shares high homology with the members of the lysR family acting as a transcriptional activator which have both of a H-T-H motif at the N-terminal region and substrate binding domain at the C-terminal region. Therefore, the orf3 is predicted as a transcriptional activator of pqrA and is designated as pqrR1. Orf4 shows homology with the periplasmic substrate-binding protein of amino acid ABC transporter. Orf5 which of incomplete sequences without stop codon revealed the homology with the permeases protein of amino acid ABC transporter.

Molecular Cloning of Chicken Major Histocompatibility Complex Class II Molecules

  • Sung, Aree-Moon
    • Toxicological Research
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    • v.8 no.2
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    • pp.331-342
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    • 1992
  • The chicken major histocompatibility complex (MHC), the B complex, is beginning to be analyzed at the DNA level. Inbred lines of chickens have been reported to possess 3~5 MHC class II genes. To further analyzed the molecular structure of the chicken MHC class II genes, cDNA clones coding for chicken MHC class II (B-L) ${\beta}$ chain molecules were isolated from chicken spleen and liver. Tissue-specific transcription of B-L ${\beta}$genes was studied by Northern blot analysis. A high level of expression was detected for spleen poly(A)$^+$ RNA whereas a faint signal was detected for liver poly(A)$^+$ RNA. Twenty-nine cDNA clones were isolated from the spleen and eight cDNA clones were isolated from the liver. Based on restriction maps, most clones could be clustered into one family of genes. Four cDNA clones were sequenced (S7, S10 and S19 from the spleen and L1, which was identical to S19, from the liver). Complete amino acid sequences of B-L ${\beta}$ chain molecules were predicated from the nucleotide sequences of the cDNA clones. Although both the nature and the location of the conserved residues were similar in chicken and mammalian sequences, some species-specific differences were found, suggesting that the structures of the B-L molecules are similar, but not identical to their mammalian counterparts.

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Optimization of artificial cultivation of Tremella fuciformis in closed culture bottle (흰목이버섯 대량생산을 위한 용기내 재배 최적화 연구)

  • Choi, Sung Woo;Chang, Hyun-You;Yoon, Jeong Weon;Lee, Chan
    • Journal of Mushroom
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    • v.6 no.1
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    • pp.20-26
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    • 2008
  • The stromatal forms of T. fuciformis and the mycelia of Hypoxylon sp. were collected. The DNA sequence in the ITS region of the 5.8S ribosomal genes of isolated strain KG103 was very similar to that of T. fuciformis AF042409 with a homology of over 98% in the EMBL/GenBank database through BLAST searching. A second isolate, No KG201, one of the symbiotic strains for cultivating T. fuciformis also exhibited high homology with Annulohhypoxylon stygium AJ390406. Potato Dextrose Medium exhibited the best mycelial growth of 14 mm/14 days and 85 mm/14 days for T. fuciformis and its symbiotic fungi, respectively. Optimum culture conditions for the micelial growth were pH 5 at $25^{\circ}C$. For the optimization of artificial cultivation of T. fuciformis in bottle with sawdust medium, several conditions such as type of sawdust, supplements, pH, moisture content, and incubation temperature were investigated. T. fuciformis and symbiotic fungi showed fast mycelial growth on corn cob media (77 and 52%) followed by oak tree sawdust and cotton seed meal. The optimal temperature for mycelial growth of T. fuciformis and symbiotic fungi on corn cob media was $25^{\circ}C$ at 55% of moisture content.

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