• Title/Summary/Keyword: group-specific sequences

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Phylogenetic Relationships among Some Bumblebees (Hymenoptera: Apidae) Common in Korea Inferred from Mitochondrial 16S rRNA Sequences

  • Yoon, Hyung-Joo;Lee, Myeong-Lyeol;Kim, Sam-Eun;Lee, Sang-Beom;Kim, Iksoo;Bae, Jin-Sik;Jin, Byung-Rae;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.1
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    • pp.51-59
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    • 2004
  • Phylogenetic relationships were analyzed among bumblebees using a portion of mitochondrial (mt) 16S ribosomal RNA (16S rRNA). Eight species of true bumblebees and one species of cuckoo bumblebee (Bombini, Apidae), collected from Korea were included in the analysis. Also, one species of true bumblebee imported from several foreign countries for pollination was included. The length of mt 16S rRNA sequence ranged from 496 bp to 508 bp and sequence divergence ranged from 1.4% (7 bp) to 15.49% (77bp). As expected, a high A+T content was observed (78.5% on average). According to the phylogeny tree derived from parsimony and maximum likelihood analysis, a monphyletic Bombus species, excluding a single cuckoo bumblebee, Psithyrus coreanus, was obtained, but the bootstrap estimate at the node supporting the monophyletic group was very weak (40% or 46%), suggesting a very close relationship of the cuckoo bumblebee to the true bumblebee. Within Bombus species belonging to identical subgenera subgeneric specific clustering was formed with high bootstrap values, implying validity of the subgeneric names of each species: Pyrobombus for B. ardens and B. modeatus; Megabombus for B. consobrinus wittenburgi and B. koreanus; and Bombus s. str. for B. ignitus, B. hypocrita sapporoensis, and B. terrestris.

Utility of Selected Non-coding Chloroplast DNA Sequences for Lineage Assessment of Musa Interspecific Hybrids

  • Swangpol, Sasivimon;Volkaert, Hugo;Sotto, Rachel C.;Seelanan, Tosak
    • BMB Reports
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    • v.40 no.4
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    • pp.577-587
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    • 2007
  • Single-copy chloroplast loci are used widely to infer phylogenetic relationship at different taxonomic levels among various groups of plants. To test the utility of chloroplast loci and to provide additional data applicable to hybrid evolution in Musa, we sequenced two introns, rpl16 and ndhA, and two intergenic spacers, psaA-ycf3 and petA-psbJ-psbL-psbF and combined these data. Using these four regions, Musa acuminata Cola(A)- and M. balbisiana Colla (B)-containing genomes were clearly distinguished. Some triploid interspecific hybrids contain A-type chloroplasts (the AAB/ABB) while others contain B-type chloroplasts (the BBA/BBB). The chloroplasts of all cultivars in 'Namwa' (BBA) group came from the same wild maternal origin, but the specific parents are still unrevealed. Though, average sequence divergences in each region were little (less than 2%), we propose that petA-psbJ intergenic spacer could be developed for diversity assessment within each genome. This segment contains three single nucleotide polymorphisms (SNPs) and two indels which could distinguish diversity within A genome whereas this same region also contains one SNP and an indel which could categorize B genome. However, an inverted repeat region which could form hairpin structure was detected in this spacer and thus was omitted from the analyses due to their incongruence to other regions. Until thoroughly identified in other members of Musaceae and Zingiberales clade, utility of this inverted repeat as phylogenetic marker in these taxa are cautioned.

PCR-based identification of uncultural bacterium from malformed Agaricus blazei (신령버섯의 기형으로부터 배양불능세균의 PCR 검정)

  • Shin, Pyung-Gyun;Park, Yun-Jung;Yoo, Young-Bok;Kong, Won-Sik;Jang, Kab-Yeul;Oh, Se-Jong;Lee, Keum-Hee
    • Journal of Mushroom
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    • v.8 no.4
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    • pp.157-160
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    • 2010
  • Agaricus blazei Murill is a important medicinal mushroom for a powerful immune system builder and tonic. Currently, it is known about a new disease phenomenon that appears to be occurring on a number of mushroom farms. We described a straightforward approach in which molecular methods was used to survey the presence of potentially endo- and epiphytic bacteria infected with the Agaricus blazei. The 16S rDNA was amplified with universal eubacterial primers directly from pure cultures of Agaricus blazei mycelium and fruit body. The 16S rDNA sequences were almost identical (96 to 97% similarity), and phylogenetic analysis placed them in a single unique rRNA branch belong to the uncultural bacterium phylogroup. PCR detection of uncultural bacterium in the malformed tissues of Agaricus blazei were carried out by using 16S rRNA sequenced specific probe. It was strongly amplified at the malformed pileus region of fruit body and also spore print was impossible.

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Effect of Titanium Surface Treatments Bond Strength and Cytotoxicity in Titanium-Porcelain System

  • Chung, In-Sung;Kim, Chi-Young;Choi, Sung-Min
    • Biomedical Science Letters
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    • v.14 no.2
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    • pp.105-113
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    • 2008
  • The objective of this study was to evaluate the influence of surface modifications on the bonding characteristics and cytotoxicity of specific titanium porcelain bonded to milling titanium and cast titanium. Milling titanium and cast titanium samples were divided into 8 test groups. These groups are as follow: i) sandblasted with particles of different size of $220{\mu}m\;and\;50{\mu}m$, ii) different sequences of sandblasting treatment and etching treatment, iii) etched with different etching solutions, and iv) preheated or not. The surface characteristics of specimens were characterized by the test of mean roughness of surface and SEM. The bond strength of titanium-ceramic systems was measured by using three-point bending test and SEM. The results show that the mean roughness of surface of sample sandblasted with $220{\mu}m$ aluminum oxide increased and bond strength were higher than sample sandblasted with $50{\mu}m$ aluminum oxide. The mean roughness of surface decreased, but the bond strength increased when the samples sandblasted with $220{\mu}m$ aluminum oxide were preheated. The sample sandblasted with $220{\mu}m$ aluminum oxide after oxidized with occupational corrosive agent I (50% NaOH, 10% $CuSO_4{\cdot}5H_2O$) and II (35% $HNO_3$, 5% HF) showed higher bond strength than sample oxidized with 30% $HNO_3$ after sandblasted with $220{\mu}m$ aluminum oxide. Group NaCuNF220SP (milling Ti: 35.3985 MPa, casting Ti: 37.2306 MPa) which was treated with occupational corrosive agent I (50% NaOH, 10% $CuSO_4{\cdot}5H_2O$) and II (35% $HNO_3$, 5% HF), followed by sandblasting with $220{\mu}m$ aluminum oxide and preheating at $750^{\circ}C$ for 1 hour showed the highest bond strength and significant differences (P<0.05). The method for modifying surface of titanium showed excellent stability of cells.

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Microbial β-Galactosidase of Pediococcus pentosaceus ID-7: Isolation, Cloning, and Molecular Characterization

  • Lee, Ji-Yeong;Kwak, Mi-Sun;Roh, Jong-Bok;Kim, Kwang;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.598-609
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    • 2017
  • Pediococcus pentosaceus ID-7 was isolated from kimchi, a Korean fermented food, and it showed high activity for lactose hydrolysis. The ${\beta}$-galactosidase of P. pentosaceus ID-7 belongs to the GH2 group, which is composed of two distinct proteins. The heterodimeric LacLM type of ${\beta}$-galactosidase found in P. pentosaceus ID-7 consists of two genes partially overlapped, lacL and lacM encoding LacL (72.2 kDa) and LacM (35.4 kDa). In this study, Escherichia coli MM294 was used for the production of LacL, LacM, and LacLM. These three types of recombinant proteins were expressed, purified, and characterized. The specific activities of LacLM and LacL were 339 and 31 U/mg, respectively. However, activity was not detected with LacM alone. The optimal pH of LacLM and LacL was pH 7.5 and pH 7.0, and the optimal temperature of LacLM and LacL was $40^{\circ}C$ and $50^{\circ}C$, respectively. The optimal temperature changes indicate that LacLM is able to achieve higher activity at a relatively lower temperature. LacLM was strongly activated by $Mg^{2+}$, $Mn^{2+}$, and $Zn^{2+}$, which was not true for LacL. Consistent with this, EDTA strongly inactivated LacLM and LacL, but the presence of reducing agents did not dramatically alter the activity. Taken together, multiple alignment of amino acid sequences and phylogenetic analysis results of LacL and LacM of P. pentosaceus ID-7 suggest the evolution of LacL into LacLM and that the use of divalent metal ions results in higher activity.

Lipase Diversity in Glacier Soil Based on Analysis of Metagenomic DNA Fragments and Cell Culture

  • Zhang, Yuhong;Shi, Pengjun;Liu, Wanli;Meng, Kun;Bai, Yingguo;Wang, Guozeng;Zhan, Zhichun;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.888-897
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    • 2009
  • Lipase diversity in glacier soil was assessed by culture-independent metagenomic DNA fragment screening and confirmed by cell culture experiments. A set of degenerate PCR primers specific for lipases of the hormone-sensitive lipase family was designed based on conserved motifs and used to directly PCR amplify metagenomic DNA from glacier soil. These products were used to construct a lipase fragment clone library. Among the 300 clones sequenced for the analysis, 201 clones encoding partiallipases shared 51-82% identity to known lipases in GenBank. Based on a phylogenetic analysis, five divergent clusters were established, one of which may represent a previously unidentified lipase subfamily. In the culture study, 11 lipase-producing bacteria were selectively isolated and characterized by 16S rDNA sequences. Using the above-mentioned degenerate primers, seven lipase gene fragments were cloned, but not all of them could be accounted for by the clones in the library. Two full-length lipase genes obtained by TAIL-PCR were expressed in Pichia pastoris and characterized. Both were authentic lipases with optimum temperatures of ${\le}40^{\circ}C$. Our study indicates the abundant lipase diversity in glacier soil as well as the feasibility of sequence-based screening in discovering new lipase genes from complex environmental samples.

Survey of Proper Primers and Genetic Analysis of Korean Ginseng (Panax ginseng C.A. Meyer) Variants using the RAPD Technique (Randomly Amplified Polymorphic DNA (RAPD) 기술을 이용한 고려인삼의 유전분석을 위한 Primer 선발 및 변종별 비교)

  • 임용표;신최순;이석종;윤영남;조재성
    • Journal of Ginseng Research
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    • v.17 no.2
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    • pp.153-158
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    • 1993
  • The study was carried out for comparison of variants and development of genetic markers using Randomly Amplified Polymorphic D사A (RAPD) analysis method. The ginseng variants used were as follows: Chungkyung-Chong, Hwangskoog-Chong, KG101 selected by the pureline selection method, and 6 kinds of Jakyung-Chong strains Uinjakyung, Jakyung-Chong 81783, Jakyung-Chong 847913, Jaky tong-Chong 79742, Jinjakyung of USSR, and Mimaki of Japan). Four of 10 RAPD primers showed the distinctive polymorphism among 9 ginseng variants and lines, and were selected for more detailed polymorphic analysis. The sequences of 4 selected primers were TGCCGAGCTG (Primer#2), AATCGGGCTG (#4), GAAACGGGTG (U7), and GTGACGTAGG (#8). All primers produced several common bands among the strains. However, when primer # 2 was applied, the electrophoregram showed the specific band at 1.8 kb region in Chungkyung-Chong, Hwangskoog- chong, and KG101, and 1 kb in the Jakyung-Chong 847913. In primer #4, 1.1 kb band was shown in Chungkyung-Chong, Hwangskoog-Chong, KG101, and Jakyung-Chong 79742. In primer # 7, 700 bp band was appeared in Jakyung-Chong 81783 and Jinjakyung of USSR In primer # 8, 800 bp band was observed only in Mimaki, comparing to another strains. When Similarity Index (SI) was calculated, Chungkyung-Chong and Hwngskoog-Chong, and Jakyung- chong 81783 and Jinjakyung of USSR showed the most close SI, 0.11 and 0.08, respectively. The data of KG101, which showed the SI of 0.13 with the group of Chungkyung-Chong and Hwangskoog-Chong, coincided with the fact that it was released from Hwangskoog-Chong by breeding process. The data of Jakyung strains indicated the significant variation among the strains. From these results, RAPD analysis method could be succesively applied to the classification and genetic analysis for breeding of Korean ginseng.

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Structure and Characteristics of Bacterial Community on Biofilm of Food Wastewater Treatment System in Winter (식품폐수 처리 공정용 생물막의 겨울철 세균군집 구조와 특성)

  • Lee, Dong-Geun;Yoo, Ki-Hwan;Park, Seong-Joo
    • Journal of Environmental Health Sciences
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    • v.37 no.2
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    • pp.124-132
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    • 2011
  • Biofilm and aeration tank of pilot and full RABC (rotating activated Bacillus contactor) plant were analyzed to characterize and determine bacterial community structure in food wastewater treatment system at winter. Concentration of heterotrophic bacteria and Bacillus group was $10^7$ and $10^5$ CFU/ml, respectively, at biofilm of pilot-plant while others represented $10^6$ and $10^4$ CFU/ml, respectively. Five and eight phyla were detected at biofilm of pilot- and full-plant, respectively, by 16S rDNA sequencing. Biofilm of pilot-plant was dominated by ${\beta}$-Proteobacteria (38.8%), ${\gamma}$-Proteobacteria (22.4%), and Bacteroidetes (12.2%), and the most dominant genus was Zoogloeae genus (22.4%). Candidate division TM7 (12.5%) was only detected at biofilm of full-plant and it was dominated by Bacteroidetes (33.3%), ${\gamma}$-Proteobacteria (29.2%), and ${\beta}$-Proteobacteria (20.8%). Clostridium genus specific primer set enabled to detect the sequences of Clostridium genus. These suggested that anaerobic and aerobic bacteria were coexisted even from the initial period of biofilm formation and ${\beta}$-Proteobacteria, ${\gamma}$-Proteobacteria and Bacteroidetes were major phyla in biofilm of food wastewater treatment system at winter.

Molecular Analyses of the Metallothionein Gene Family in Rice (Oryza sativa L.)

  • Zhou, Gongke;Xu, Yufeng;Li, Ji;Yang, Lingyan;Liu, Jin-Yuan
    • BMB Reports
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    • v.39 no.5
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    • pp.595-606
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    • 2006
  • Metallothioneins are a group of low molecular mass and cysteine-rich metal-binding proteins, ubiquitously found in most living organisms. They play an important role in maintaining intracellular metal homeostasis, eliminating metal toxification and protecting against intracellular oxidative damages. Analysis of complete rice genome sequences revealed eleven genes encoding putative metallothionein (OsMT), indicating that OsMTs constitute a small gene family in rice. Expression profiling revealed that each member of the OsMT gene family differs not only in sequence but also in their tissue expression patterns, suggesting that these isoforms may have different functions they perform in specific tissues. On the basis of OsMT structural and phylogenetic analysis, the OsMT family was classified as two classes and class I was subdivided into four types. Additionally, in this paper we also present a complete overview of this family, describing the gene structure, genome localization, upstream regulatory element, and exon/intron organization of each member in order to provide valuable insight into this OsMT gene family.

Community Structure of Bacteria Associated with Two Marine Sponges from Jeju Island Based on 16S rDNA-DGGE Profiles (16S rDNA-DGGE를 이용한 2종의 제주도 해양 해면의 공생세균의 군집 구조)

  • Park, Jin-Sook;Sim, Chung-Ja;An, Kwang-Deuk
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.170-176
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    • 2009
  • Culture-independent 16S rDNA-DGGE profiling and phylogenetic analysis were used to examine the predominant bacterial communities associated with the two sponges, Dictyonella sp. and Spirastrella abata from Jeju island. The culture-independent approach involved extraction of total bacterial DNA, PCR amplification of the 16S ribosomal DNA using primer pair 341f-GC and 518r, and separation of the amplicons on a denaturing gradient gel. Denaturing gradient gel electrophoresis banding patterns indicated 8 and 7 bands from the two sponge species, Dictyonella sp. and Spirastrella abata, respectively. There were not common major bands in two different sponges. Comparative sequence analysis of variable DGGE bands revealed from 93% to 98% similarity to the known published sequences. The dominant bacterial group of Dictyonella sp. belonged to uncultured Gammaproteobacteria, while, that of Spirastrella abata belonged to uncultured Alphaproeobacteria and Firmicutes. DGGE analysis indicated predominant communities of the sponge-associated bacteria differ in the two sponges from the same geographical location. This result revealed that bacterial community profiles of the sponges were host species-specific.