• Title/Summary/Keyword: DNA homology

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BCR (Brown Color Repressor) gene isolation related to mycelial browning of Lentinus edodes (표고균사 갈변과 관련된 BCR (Brown Color Repressor) 유전자 분리)

  • Kim, Young-Ho;Park, Soo-Cheol;Jhune, Chang-Sung;You, Chang-Hyun;Sung, Jae-Mo;Kong, Won-Sik
    • Journal of Mushroom
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    • v.10 no.3
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    • pp.120-128
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    • 2012
  • Recently sawdust cultivation of Shiitake mushroom (Lentinula edodes) is increasing. It is important to make mycelia to be brown on the substrate surface. This browned surface in sawdust cultivation plays an important role like as artificial bark of the oak log, which protects the other pests and suppresses water evaporation in the substrate. In order to isolate genes which related to brown color formation, differential display method was used. Two cDNA fragments obtained by DD-PCR were 1.2 and 1.6kb and these were expressed in white colored mycelia from L. edodes, but not brown colored mycelia. Partial sequencing of these cDNA fragments showed that the 1.6kb cDNA had 100% identity with the microsatellites gene from Dugenia polichroa. However, the other 1.2kb cDNA fragment had poly T tail on 3' region of partial open reading frame on 5' region. The new primer designed based on the sequence of 1.2kb cDNA was constructed. RT-PCR analysis using the newly designed 0.12kb cDNA specific primer showed that the gene was only expressed in white color mycelia, but not in brown color mycelia. Sequence analysis of 5' region of this 1.2kb cDNA revealed that this gene contained partial open reading frame consisted of 110 amino acid. Homology search using DNASIS database showed that this gene had high sequence homology of 66.7% in DNA level and 69.2 % in amino acid level with dTDP-glucose 4,6-dehydratases gene from Arabidopsis thaliata. The dTDP-glucose 4,6-dehydratases gene was known to be function to have tolerance with oxidation stress. These results strongly suggest that this gene isolated from white mycelia of L. edodes might have a function of repressor against mycelia browning. Therefore I designated this gene as BCR (Brown Color Repressor) gene.

Nucleotide and Deduced Amino Acid Sequences of Rat Myosin Binding Protein H (MyBP-H)

  • Jung, Jae-Hoon;Oh, Ji-Hyun;Lee, Kyung-Lim
    • Archives of Pharmacal Research
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    • v.21 no.6
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    • pp.712-717
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    • 1998
  • The complete nucleotide sequence of the cDNA clone encoding rat skeletal muscle myosin- binding protein H (MyBP-H) was determined and amino acid sequence was deduced from the nucleotide sequence (GenBank accession number AF077338). The full-length cDNA of 1782 base pairs(bp) contains a single open reading frame of 1454 bp encoding a rat MyBP-H protein of the predicted molecular mass 52.7kDa and includes the common consensus 1CA__TG' protein binding motif. The cDNA sequence of rat MyBP-H show 92%, 84% and 41% homology with those of mouse, human and chicken, respectively. The protein contains tandem internal motifs array (-FN III-Ig C2-FN III- Ig C2-) in the C-terminal region which resembles to the immunoglobulin superfamily C2 and fibronectin type III motifs. The amino acid sequence of the C-terminal Ig C2 was highly conserved among MyBPs family and other thick filament binding proteins, suggesting that the C-terminal Ig C2 might play an important role in its function. All proteins belonging to MyBP-H member contains `RKPS` sequence which is assumed to be cAMP- and cGMP-dependent protein kinase A phosphorylation site. Computer analysis of the primary sequence of rat MyBP-H predicted 11 protein kinase C (PKC)phosphorylation site, 7 casein kinase II (CK2) phosphorylation site and 4N-myristoylation site.

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Cloning and Characterization of Novel Soluble Acid Invertase Which is Responsible to JA, ABA and GA During Tip Growth of Pea Seedlings (Pisum sativum)

  • Kim, Dong-Giun;Zhang, Jiesheng
    • Korean Journal of Environmental Biology
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    • v.27 no.4
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    • pp.406-413
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    • 2009
  • The enzyme invertase contributes to sugar unloading, pathogen defense, differentiation and development in plants. We cloned the complete cDNA of a soluble acid invertase from pea seedlings (Pisum sativum) via RT-PCR and the rapid amplification of the cDNA end (RACE) technique. The full-length cDNA of the soluble pea invertase comprised 2237 bp and contained a complete open reading frame encoding 647 amino acids. The deduced amino acid sequence showed high homology to soluble acid invertases from various plants. Northern blot analysis demonstrated the soluble acid invertase gene of P. sativum was strongly expressed in sink organs such as shoot tips and root tips, and induced by abscisic acid, gibberellic acid and jasmonic acid in shoots. Especially, gibberellic acid enhanced the gene expression of the soluble acid invertase in a time-dependent manner. This study presents that the gene expression patterns of a soluble acid invertase from pea are strongly consistent with the suggestion that individual invertase gene product has different functions in the growing plant.

Population Analysis of Korean and Japanese Toxic Alexandrium catenella Using PCR Targeting the Area Downstream of the Chloroplast PsbA Gene

  • Kim Choong-Jae;Kim Chang-Hoon;Sako Yoshihiko
    • Fisheries and Aquatic Sciences
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    • v.7 no.3
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    • pp.130-135
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    • 2004
  • The marine dinoflagellate genus Alexandrium, which produces PSP toxins, has a global distribution. As human-assisted dispersal of the species has been suggested, it is important to develop molecular tools to trace the dispersal pathway. To screen population-specific DNA sequences that differentiate Korean and Japanese A. catenella, we targeted the area downstream of the chloroplast psbA gene using PCR with population-specific DNA primers followed by RFLP (restriction fragment length polymorphism) analysis and sequencing. The RFLP patterns of the PCR products divided Korean and Japanese A. catenella regional isolates into three types: Korean, Japanese, and type CMC3, isolated from Korea. We sequenced the PCR products, but found no similar gene in a homology search. The molecular phylogeny inferred from the sequences separated the Korean and Japanese A. catenella strains, as did the RFLP patterns. However, the Japanese isolates included two slightly different sequences (types J and K), while the Korean sequence was the same as the Japanese K type. In addition, a unique sequence was found in the Korean strains CMC2 and CMC3. Population-specific PCR amplification with Japanese A. catenella type-specific PCR primers designed from the type J sequence yielded PCR products for Japanese strains only, showing that the unknown gene can be used for a population analysis of Korean and Japanese A. catenella.

Characterization and cDNA Cloning of a Defensin-Like Peptide, Harmoniasin, from Harmonia axyridis

  • Kim, In-Woo;Lee, Joon-Ha;Park, Ha-Yan;Kwon, Young-Nam;Yun, Eun-Young;Nam, Sung-Hee;Kim, Seong-Ryul;Ahn, Mi-Young;Hwang, Jae Sam
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1588-1590
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    • 2012
  • We compared the mRNA expression profile of the Harmonia axyridis larvae that were either untreated or treated with LPS. The extracted mRNAs were subjected to ACP RT-PCR analysis using a combination of arbitrary primers and oligo (dT) primer. Among the 47 DEGs differentially expressed, we identified a cDNA showing homology with defensin-like antibacterial peptide. The cDNA showed a putative 32-residue signal sequence and a 50-residue mature peptide named harmoniasin. We also investigated the antibacterial activity of the harmoniasin analog, which exhibited potent antibacterial activities against Gramnegative and -positive bacteria strains and it also evidenced no hemolytic activity.

Characterization and identification of heterotrophic nitrifying bacteria isolated from composting soil (퇴비로부터 분리한 종속영양 질화세균의 동정 및 특징에 관한 연구)

  • Kim, Young-Jun;Lee, Gun-Young;Kim, Jin-Yong
    • Journal of the Korea Organic Resources Recycling Association
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    • v.13 no.2
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    • pp.121-127
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    • 2005
  • A heterotrophic nitrifying bacterium was isolated from the compost and analyzed for its characteristics. This bacterium was found to be a Gram positive rod, catalase positive, and motile. Nitrite production was detected on the ammonium acetate medium through the violet color formation. BBL test showed that this strain has high homology with Bacillus strains. Phylogenetic analysis using 16S rDNA revealed that the bacterium has 94% of similarity with Mycobacterium smegmatis strain.

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Molecular cloning of a rhoptry protein (ROP6) secreted from Toxoplasma gondii

  • Ahn Hye-Jin;Kim Seh-Ra;Nam Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.44 no.3
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    • pp.251-254
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    • 2006
  • Monoclonal antibody (mAb) Tg786 against Toxoplasma gondii has been found to detect a 42-kDa rhoptry protein (ROP6) which showed protease activity and host cell binding characteristics after secretion. Using the mAb, a colony containing a 3'-UTR was probed in a T. gondii cDNA expression library. A full length cDNA sequence of the rhoptry protein was completed after 5'-RACE, which consisted of 1,908 bp with a 1,443 bp ORF. The deduced amino acid sequence of ROP6 consisted of a polypeptide of 480 amino acids without significant homology to any other known proteins. This sequence contains an amino terminal stop transfer sequence downstream of a short neutral sequence, hydrophilic middle sequence, and hydrophobic carboxy terminus. It is suggested that the ROP6 is inserted into the rhoptry membrane with both N- and C-termini.

Genetic characterization of Phellinus baumii PMO-P4 by analyzing restriction fragment length polymorphisms of nuclear ribosomal DNA internal transcribed spacers (ITS) (Ribosomal DNA의 ITS부위에 대한 RFLP 분석에 의한 Phellinus baumii PMO-P4의 유전학적 특성)

  • Chang, Yun-Hee;Kim, Tae-Rack;Kim, Hyun-Su;Yeo, Ik-Hyun;Lee, Sang-Youn;Ha, Hyo-Cheol
    • Journal of Mushroom
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    • v.4 no.2
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    • pp.43-47
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    • 2006
  • PMO-P4, being cultivated as "Sanghwang" in Korea, was proved to be P. baumii based on ITS (internal transcribed spacer) sequencing and RFLP (Restriction Fragment Length Polymorphism) patterns along with some Phellinus species including P. linteus. The similaraty of ITS sequencing between PMO-P4 and other Phellinus species was given the range of 48.6%~72.2%, showing the highest homology from P. linteus and the lowest from P. gilvus.

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Isolation and characterization of Phytochrome B gene in Poplar (포플러의 Phytochrome B 유전자 분리 및 특성구명)

  • Kang, Hoduck;Lee, Keum-Young;Kang, Sang-Gu;Bae, Han-hong
    • Journal of Korean Society of Forest Science
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    • v.94 no.4 s.161
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    • pp.236-242
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    • 2005
  • Phytochrome B (PhyB) gene, which is a photoreceptor that controls plant growth under various light conditions, was cloned from Chinese hybrid poplar 'Soohang 1'. Nucleotide sequence and deduced amino acid sequences PhyB cDNA of 'Soohang' is consisted with 3,456 nucleotides and 1,156 amino acids. The cloned PhyB fragment showed 98% homology of amino acid sequences with Populus balsamifera PhyB1. According to Northern blot analysis. PhyB was up-regulated by light, while PhyB transcript was not detected under dark condition. According to this study, the cloned PhyB is induced by light and functions as photoreceptor.

Studies on the cloning gp50 and gp63 genes of Pseudorabies virus(Shope strain) (Pseudorabies virus의 gp50과 gp63 유전자 클로닝에 관한 연구)

  • Kweon, Chang-hee;Song, Jae-young;Kim, Byoung-han;Lee, Jung-bok;Lee, jae-chin;An, Soo-hwan;Lee, Yong-soon;Susumu, Maeda
    • Korean Journal of Veterinary Research
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    • v.31 no.3
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    • pp.311-318
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    • 1991
  • The DNA fragment representing for Pseudorabies gp50 and gp60(Shope) was cloned by recombinant techniques. The viral DNA was extracted from the infected cells and digested with Bam HI. The 6.8 Kb of Bam HI fragment was isolated from agarose gel and further digested with Nde I followed by Klenow treatment. The blunt ended 4.9Kb fragment was cloned into pTZ18R plasmid vector. The upstream region of gp50 was further manipulated to remove its 5' promoter region and create EcoRl site for possible eukaryotic expression system. The result of partial sequencing of cloned DNA indicated that Shope strain showed 95% homology with gp50 of Rice strain.

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