• Title/Summary/Keyword: Environmental clone

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Hypersensitivity of Somatic Mutations and Mitotic Recombinations Induced by Mutagens in Transgenic Drosophila bearing Rat DNA Polymerase $\beta$ (Rat의 DNA Polymerase$\beta$ cDNA가 도입된 Transgenic Drosophila의 체세포 돌연변이 유발에 관한 연구)

  • 최영현;유미애;이원호
    • Environmental Mutagens and Carcinogens
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    • v.15 no.2
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    • pp.100-105
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    • 1995
  • The effects of DNA polymerase $\beta$ on the somatic chromosome mutations and mitotic recombinations were investigated using the transgenic Drosophila beating chimetic gene consisting of a promoter region of Drosophila actin 5C gene and rat DNA polymerase $\beta$. For detecting the somatic chromosome mutations and mitotic recombinations, the heterozygous (mwh/+) strains possessing or lacking transgene poi 13 were used. The spontaneous frequency of small mwh spots, due to deletion or nondisjunction etc., in the non-transgenic w strain and the transgenic p[pol $\beta$]-130 strain was 0.351 and 0.606, respectively. The spontaneous frequency (0.063) of large mwh spots, arises mostly from somatic recombination between the centromere and the locus mwh, in the transgenic p[pol $\beta$]-130 strain was about three times higher than that (0.021) of the non-transgenic w strain. The mutant clone frequencies of small and large mwh spots induced by N-methyl-N'-nitro-N-nitrosoguanidine and ethyl methanesulfonate in the transformant p[pol $\beta$]-130 were higher than those in the host strain w. The present results suggest that rat DNA polymerase $\beta$ participate at least in the somatic chromosome mutations and mitotic recombination processes.

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Determination of Complete Genome Sequence of Korean Isolate of Potato virus X

  • Choi, Sun-Hee;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • v.24 no.3
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    • pp.361-364
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    • 2008
  • The complete nucleotide sequences of a Korean isolate of Potato virus X(PVX-Kr) has been determined. Full-length cDNA of PVX-Kr has been directly amplified by long template reverse transcription and polymerase chain reaction(RT-PCR) using virus specific 5'-end primer and 3'-end primer, and then constructed in a plasmid vector. Consecutive subclones of a full-length cDNA clone were constructed to identify whole genome sequence of the virus. Total nucleotide sequences of genome of PVX-Kr were 6,435 excluding one adenine at poly A tail, and genome organization was identical with that of typical PVX species. Comparison of whole genome sequence of PVX-Kr with those of European and South American isolates showed 95.4-96.8% and 77.4-77.9%, in nucleotide similarity, respectively. Sequenced PVX-Kr in this study and twelve isolates already reported could be divided into two subgroups in phylogeny based on their complete nucleotide sequences. Phylogenetic tree analysis demonstrated that PVX-Kr was clustered with European and Asian isolates(Taiwan, os, bs, Kr, S, X3, UK3, ROTH1, Tula) in the same subgroup and South American isolates(CP, CP2, CP4, HB) were clustered in the other subgroup.

Diversity of Nitrifying and Denitrifying Bacteria in SMMIAR Process (완전침지형 회전매체공정 내 질산화 및 탈질 관련 미생물의 군집 분포)

  • Quan, Zhe-Xue;Lim, Bong-Su;Kang, Ho;Yoon, Kyung-Yo;Yoon, Yeo-Gyo
    • Journal of Korean Society on Water Environment
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    • v.22 no.6
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    • pp.1014-1021
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    • 2006
  • SMMIAR (Submerged Moving Media Intermittent Aeration Reactor) Process is a very efficient system which remove ammonia to nitrogen gas in one reactor. In this study, we determined the diversity of ammonia oxidizing bacteria and denitrifying bacteria using specific PCR amplification and the clone library construction. An ammonia monooxygenase gene(amoA) was analyzed to investigate the diversity of nitrifiers. Most of amoA gene fragments (27/29, 93%) were same types and they are very similar (>99%) to the sequences of Nitrosomonas europaea and other clones isolated from anoxic ammonia oxidizing reactors. ANAMMOX related bacteria have not determined by specific PCR amplification. A nitrite reductase gene(nirK) was analyzed to investigate the diversity of denitrifying bacteria. About half (9/20, 45%) of denitrifiers were clustered with Rhodobacter and most of others were clustered with Mesorhizobium (6/20, 30%) and Rhizobium (3/20, 15%). All of these nirK gene clones were clustered in alpha-Proteobacteria and this result is coincide with other system which also operate nitrification and denitrification in one reactor. The molecular monitoring of the population of nitrifiers and denitrifiers would be helpful for the system stabilization and scale-up.

Remarkable Bacterial Diversity in the Tidal Flat Sediment as Revealed by 16S rDNA Analysis

  • Chun, Jong-Sik;Kim, Bong-Soo;Oh, Huyn-Myung;Kang, Ho-Jeong;Park, Seok-Soon
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.205-211
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    • 2004
  • A 16S rDNA clone library was generated to investigate the bacterial diversity in tidal flat sediment in Ganghwa Island, Republic of Korea. A total of 103 clones were sequenced and analyzed by comprehensive phylogenetic analyses. No clones were identical to any of known 16S rRNA sequences in public databases. Sequenced clones fell into thirteen lineages of the domain Bacteria: the alpha, beta, gamma, delta, and epsilon Proteobacteria, Actinobacteria, CFB group, Chloroflexi, Acidobacteria, Planctomycetes, Verrucomicrobia, and known uncultured candidate divisions (OP11, BRC1, KSB1, and WS1). Two clones were not associated with any known bacterial divisions. The majority of clones belonged to the gamma and delta Proteobacteria (46.7%). Clones of Actinobacteria were distantly related to known taxa. It is evident from 16S rDNA-based community analysis that the bacterial community in tidal flat sediment is remarkably diverse and unique among other marine environments examined so far.

Isolation and Characterization of Phosphorus Accumulating Microorganisms under Liquid Fertilization of Swine Slurry (액비화 과정 중 인 이용 우수미생물 분리 및 특성)

  • Lim, Joung-Soo;Cho, Sung-Back;Hwang, Ok-Hwa;Yang, Seung-Hak
    • Journal of Animal Environmental Science
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    • v.20 no.2
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    • pp.77-84
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    • 2014
  • This study was conducted to investigate the bacterial capability to accumulate phosphorus during liquid composting process of pig slurry. Storage liquid compost and pig slurry were analyzed by using MALDI-TOF technique, which showed the colonies of Acinetobacter towneri and Bacillus licheniformis. In addition, bacterial colonies were isolated under high phosphoric acid conditions using X-phosphate MOPS medium with the addition of 2 mM $K_2HPO_4$. Microbial growth was observed in high and low phosphoric conditions due to the growth of bacterial diversities in the liquid fertilizer and slurry. The colonies isolated in the high phosphoric acid medium were uncultured bacterium clone and Acinetobacter sp. were identified by analysis of 16S rRNA gene sequences. Uncultured bacterium showed higher growth rate and excellent phosphorus ability then Acinetobacter sp.. In addition to Paenibacillus sp. AEY-1 isolated from pig slurry performed excellent phosphorus utilizing capability.

Identification of Chinese Cabbage Sentrin as a Suppressor of Bax-Induced Cell Death in Yeast

  • Sawitri, Widhi Dyah;Slameto, Slameto;Sugiharto, Bambang;Kim, Kyung-Min
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.600-606
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    • 2012
  • Studies into the cell death program termed apoptosis have resulted in new information regarding how cells control and execute their own demise, including insights into the mechanism by which death-preventing factors can inhibit Bax-induced caspase activation. We investigated high temperature stress-induced cell death in Brassica rapa. Using a yeast functional screening from a Brassica rapa cDNA library, the BH5-127 EST clone encoding an apoptotic suppressor peptide was identified. However, a phylogenic tree showed that BH5-127 clusters within a clade containing SUMO-1 (Small Ubiquitin-like Modifier-1). BH5-127 was confirmed similar to have function to SUMO-1 as Fas suppression. Expression of BH5-127 showed that substantial suppression of cell death survived on SD-galactose-$Leu^-$-$Ura^-$ medium. The results suggest that BrSE ($\underline{B}$rassica rapa $\underline{S}$entrin $\underline{E}$ST, BH5-127) is one of the important regulatory proteins in programming cell death, especially in the seedling stage of Chinese cabbage.

Identification and Molecular Characterization of Parkin in Clonorchis sinensis

  • Bai, Xuelian;Kim, Tae Im;Lee, Ji-Yun;Dai, Fuhong;Hong, Sung-Jong
    • Parasites, Hosts and Diseases
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    • v.53 no.1
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    • pp.65-75
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    • 2015
  • Clonorchis sinensis habitating in the bile duct of mammals causes clonorchiasis endemic in East Asian countries. Parkin is a RING-between-RING protein and has E3-ubiquitin ligase activity catalyzing ubiquitination and degradation of substrate proteins. A cDNA clone of C. sinensis was predicted to encode a polypeptide homologous to parkin (CsParkin) including 5 domains (Ubl, RING0, RING1, IBR, and RING2). The cysteine and histidine residues binding to $Zn^{2+}$ were all conserved and participated in formation of tertiary structural RINGs. Conserved residues were also an E2-binding site in RING1 domain and a catalytic cysteine residue in the RING2 domain. Native CsParkin was determined to have an estimated molecular weight of 45.7 kDa from C. sinensis adults by immunoblotting. CsParkin revealed E3-ubiquitin ligase activity and higher expression in metacercariae than in adults. CsParkin was localized in the locomotive and male reproductive organs of C. sinensis adults, and extensively in metacercariae. Parkin has been found to participate in regulating mitochondrial function and energy metabolism in mammalian cells. From these results, it is suggested that CsParkin play roles in energy metabolism of the locomotive organs, and possibly in protein metabolism of the reproductive organs of C. sinensis.

AFLP analysis to assess genomic stability in Solanum regenerants derived from wild and cultivated species

  • Aversano, Riccardo;Di Dato, Francesco;Di Matteo, Antonio;Frusciante, Luigi;Carputo, Domenico
    • Plant Biotechnology Reports
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    • v.5 no.3
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    • pp.265-271
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    • 2011
  • The cultivated potato as well as its tuber-bearing relatives are considered model plants for cell and tissue culture, and therefore for exploiting the genetic variation induced by in vitro culture. The association between molecular stability and tissue culture in different genetic backgrounds and ploidy levels has already been explored. However, it still remains to be ascertained whether somaclonal variation differs between callus-derived chromosome-doubled and undoubled regenerants. Our research aimed at investigating, through amplified fragment length polymorphism (AFLP) markers, the genetic changes in marker-banding patterns of diploid and tetraploid regenerants obtained from one clone each of Solanum bulbocastanum Dunal and S. cardiophyllum Lindl (both 2n = 2x = 24) and tetraploids from cultivated S. tuberosum L. (2n = 4x = 48). Pairwise comparisons between the banding patterns of regenerants and parents allowed detecting considerable changes associated to in vitro culture both at diploid and tetraploid level. The percentages of polymorphic bands between diploid and tetraploid regenerants were, respectively, 57 and 69% in S. bulbocastanum and 58 and 63% in S. cardiophyllum. On average, the frequencies of lost parental fragments in regenerants were significantly higher than novel bands both in S. bulbocastanum (48 vs. 22%) and S. tuberosum (36 vs. 18%) regenerants. By contrast, in S. cardiophyllum, a similar incidence of the two events was detected (32 vs. 29%). Our results revealed that structural changes after tissue culture process strongly affected the genome of the species studied, but diploid and tetraploids regenerated plants responded equally.

Analysis of Sequence Diversity of mcyA Gene Involved in Microcystin Synthesis in Korean Reservoirs (국내 호수에서 Microcystins의 생합성에 관여하는 mcyA 유전자의 염기서열 다양성 분석)

  • Oh, Kyoung-Hee;Han, Ah-Won;Cho, Young-Cheol
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.162-168
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    • 2010
  • The sequence diversity of mcyA gene involved in synthesis of microcystins was analyzed in Microcystis spp. isolated from the Korean reservoirs and in the environmental samples taken from the Daechung, Chungju, Yongdam, Soyang, and Euam Reservoirs at the cyanobacterial blooming season. It was estimated that the sequences of mcyA gene in the isolated Microcystis spp. were much conserved when compared with those in GenBank database. A few kinds of clones were dominant in the investigated environmental samples, occupying 87 to 100% of total clones. No mcyA sequences originated from Anabaena spp. or Planktothrix spp. was found. These results indicated that microcystins are produced mainly by Microcystis spp. and the sequences of mcyA genes are much conserved in the investigated Korean reservoirs.

T-RFLP Analysis of Microbial Community Structure in Leachate from Landfill Sites (폐기물매립장 침출수내 미생물군집 구조 해석을 위한 T-RFLP의 활용)

  • Yu, Jae-Cheul;Ishigaki, Tomonori;Kamagata, Yoichi;Lee, Tae-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.4
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    • pp.369-378
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    • 2010
  • Microorganisms are key-role player for stabilization of landfill sites. In order to evaluate the availability of T-RFLP(Terminal Restriction Fragment Length Polymorphism) for monitoring microbial community variations during stabilization of landfill sites, the phylogenic diversity of microbial community in the leachate from 4 different full-scale landfills was characterized by T-RFLP based on bacterial 16S rDNA. Main population of microbial community analyzed by T-RFLP was significantly similar with that of microbial community analyzed by clone library analysis. The results of T-RFLP analysis for main population of microbial community in the leachate from landfills with different landfill structures, waste types and landfill ages showed apparently different microbial diversity and structures. Therefore, long-term monitoring of microbial community in leachate from landfill sites by using T-RFLP is expected to be available for evaluation of landfill stability.