• Title/Summary/Keyword: copy numbers

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Diversity and Abundance of Ammonia-Oxidizing Bacteria in Activated Sludge Treating Different Types of Wastewater

  • Baek, Kyung-Hwa;Park, Chul;Oh, Hee-Mock;Yoon, Byung-Dae;Kim, Hee-Sik
    • Journal of Microbiology and Biotechnology
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    • 제20권7호
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    • pp.1128-1133
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    • 2010
  • The diversity and abundance of ammonia-oxidizing bacteria (AOB) in activated sludge were compared using PCR-DGGE and real-time PCR assays. Activated sludge samples were collected from five different types of wastewater treatment plants (WWTPs) mainly treating textile, paper, food, and livestock wastewater or domestic sewage. The composition of total bacteria determined by PCR-DGGE was highly diverse between the samples, whereas the community of AOB was similar across all the investigated activated sludge. Total bacterial numbers and AOB numbers in the aerated mixed liquor were in the range of $1.8{\times}10^{10}$ to $3.8{\times}10^{12}$ and $1.7{\times}10^6$ to $2.7{\times}10^{10}$ copies/l, respectively. Activated sludge from livestock, textile, and sewage treating WWTPs contained relatively high amoA gene copies (more than $10^5$ copies/l), whereas activated sludge from food and paper WWTPs revealed a low number of the amoA gene (less than $10^3$ copies/l). The value of the amoA gene copy effectively showed the difference in composition of bacteria in different activated sludge samples and this was better than the measurement with the AOB 16S rRNA or total 16S rRNA gene. These results suggest that the quantification of the amoA gene can help monitor AOB and ammonia oxidation in WWTPs.

제주도 연안 해역의 오염지표세균의 분포 (Distribution of Indicator Bacteria in Seawater off the Coast of Jeju Island)

  • 노형진;임윤진;김아란;김남은;김영재;박노백;황지연;권문경;김도형
    • 한국수산과학회지
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    • 제51권4호
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    • pp.450-455
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    • 2018
  • We examined correlations of the density of fish farms with the distributions of indicator bacteria (Escherichia coli, fecal streptococci) and a bacterial fish pathogen (Streptococcus parauberis) off the coastline of Jeju Island. Seawater samples were collected at four coastal sites on the Island [Aewol (control), Gujwa, Pyoseon and Daejeong] in June, August and October 2016. The indicator bacteria were generally more frequently isolated from samples taken in August when water temperatures and human activities on nearby beaches were highest. Although fish farms were least common at Daejeong, the numbers of isolated fecal indicator bacteria were highest in the seawater and effluent water collected from this site. Hence, fish farms were not likely major contributors of indicator bacteria at Daejeong. We found discrepancies between the isolated bacterial counts and the predicted bacterial copy numbers deduced from our qPCR results, indicating that this pathogen may exist in a viable but non-culturable (VBNC) state in seawater. Thus, livestock wastewater and chemical fertilizer loading off Jeju Island may negatively impact seawater quality more than the effluent released from fish farms does.

상향식 바이오필터에서 상.하층의 메탄 산화 특성 비교 (Comparison of Methanotrophic Activity at Top and Bottom Layers in Up-flow Biofilters)

  • 윤정희;김정미;김지은;이다슬;조경숙
    • 한국미생물·생명공학회지
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    • 제41권2호
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    • pp.221-227
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    • 2013
  • Perlite와 tobermolite 담체를 각각 상 하단(바이오필터 A) 또는 하 상단(바이오필터 B)으로 충전한 상향식 바이오필터에서 상하단의 메탄산화 특성을 비교하였다. 각 바이오필터에서 상단과 하단 담체를 채취하여 메탄산화속도를 측정하고, 정량적 real time PCR 방법을 이용하여 메탄산화세균수를 정량분석 하였다. 혼합담체의 층적배열 차이에 따른 각 담체의 순수 메탄산화속도를 조사한 결과, biofilter A perlite 상단이 $845.16{\pm}64.78{\mu}mol{\cdot}VS^{-1}{\cdot}h^{-1}$로 tobermolite 하단 $381.85{\pm}42.00{\mu}mol{\cdot}VS^{-1}{\cdot}h^{-1}$에 비하여 상대적으로 높았다(p < 0.005). 또한, biofilter B tobermolite의 상단($601.25{\pm}37.78{\mu}mol{\cdot}VS^{-1}{\cdot}h^{-1}$)이 perlite 하단($411.07{\pm}53.02{\mu}mol{\cdot}VS^{-1}{\cdot}h^{-1}$)에 비하여 메탄산화속도가 높았다(p < 0.005). 바이오필터 A는 상단(1.27E+13 pmoA gene copy number/mg-VSS)보다는 하단(3.33E+13 pmoA gene copy number/mg-VSS) (p < 0.05)에 더 많은 메탄 산화 세균이 존재하였다. 그러나, 바이오필터 B는 상단과 하단간의 메탄 산화 세균수의 차이는 유의하지 않은 것으로 나타났다(p > 0.05). Perlite와 tobermolite로 충진된 각 담체의 순수 메탄산화속도는 상단부에 위치한 담체들이 높았음에도 불구하고 바이오필터내에서 메탄농도 감소폭이 컸던 하단부에서 메탄산화세균이 많이 존재하였다.

Perspectives of Integrative Cancer Genomics in Next Generation Sequencing Era

  • Kwon, So-Mee;Cho, Hyun-Woo;Choi, Ji-Hye;Jee, Byul-A;Jo, Yun-A;Woo, Hyun-Goo
    • Genomics & Informatics
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    • 제10권2호
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    • pp.69-73
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    • 2012
  • The explosive development of genomics technologies including microarrays and next generation sequencing (NGS) has provided comprehensive maps of cancer genomes, including the expression of mRNAs and microRNAs, DNA copy numbers, sequence variations, and epigenetic changes. These genome-wide profiles of the genetic aberrations could reveal the candidates for diagnostic and/or prognostic biomarkers as well as mechanistic insights into tumor development and progression. Recent efforts to establish the huge cancer genome compendium and integrative omics analyses, so-called "integromics", have extended our understanding on the cancer genome, showing its daunting complexity and heterogeneity. However, the challenges of the structured integration, sharing, and interpretation of the big omics data still remain to be resolved. Here, we review several issues raised in cancer omics data analysis, including NGS, focusing particularly on the study design and analysis strategies. This might be helpful to understand the current trends and strategies of the rapidly evolving cancer genomics research.

Identification of Novel Universal Housekeeping Genes by Statistical Analysis of Microarray Data

  • Lee, Se-Ram;Jo, Min-Joung;Lee, Jung-Eun;Koh, Sang-Seok;Kim, So-Youn
    • BMB Reports
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    • 제40권2호
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    • pp.226-231
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    • 2007
  • Housekeeping genes are widely used as internal controls in a variety of study types, including real time RT-PCR, microarrays, Northern analysis and RNase protection assays. However, even commonly used housekeeping genes may vary in stability depending on the cell type or disease being studied. Thus, it is necessary to identify additional housekeeping-type genes that show sample-independent stability. Here, we used statistical analysis to examine a large human microarray database, seeking genes that were stably expressed in various tissues, disease states and cell lines. We further selected genes that were expressed at different levels, because reference and target genes should be present in similar copy numbers to achieve reliable quantitative results. Real time RT-PCR amplification of three newly identified reference genes, CGI-119, CTBP1 and GOLGAl, alongside three well-known housekeeping genes, B2M, GAPD, and TUBB, confirmed that the newly identified genes were more stably expressed in individual samples with similar ranges. These results collectively suggest that statistical analysis of microarray data can be used to identify new candidate housekeeping genes showing consistent expression across tissues and diseases. Our analysis identified three novel candidate housekeeping genes (CGI-119, GOLGA1, and CTBP1) that could prove useful for normalization across a variety of RNA-based techniques.

Regulation of the expression of nhaA gene coding $Na^{+}$/$H^{+}$ antiporter A of escherichia coli

  • Seo, Sung-Yum;Lee, Seung-Heon
    • Journal of Microbiology
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    • 제33권2호
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    • pp.120-125
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    • 1995
  • .betha.-galactosidase activity of Escherichia coli cells containing operon fusion nhaA'-'lacZ was monitored to study the regulation of expression of nhaA gene under various conditions. The expression of the fusion was enhanced only by chemicals containing Na$^{+}$ or Li$^{+}$. This Na$^{+}$ or Li$^{+}$. This Na$^{+}$(Li$^{+}$)-specific enhancement of .betha.-galactosidase activity represented the increase in the rate of synthesis of .betha.-galactosidase rather than the decrease in the breakdown rate. The induction pattern was influenced by copy numbers of the gene. Induction by Na$^{+}$ or Li$^{+}$ was concentration and time dependent, reaching maximum 5-6 fold induction after 2 hours at 0.4-0.5 M for Na$^{+}$ or at 0.25-0.35 M for Li$^{+}$, Although the expression was induced at much lower concentration of Na$^{+}$ at alkaline pH values than at neutral pH in the presence of Na$^{+}$, alkaline pH itself did ot induced the expression of the fusion in the absence of Na$^{+}$. Temperature shift and growth phase of culture did not affect the level of induction.he level of induction.

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Identification and Phylogeny of the Human Endogenous Retrovirus HERV-W LTR Family in Human Brain cDNA Library and Xq21.3 Region

  • KIM, HEUI-SOO;TIMOTHY J. CRO
    • Journal of Microbiology and Biotechnology
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    • 제12권3호
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    • pp.508-513
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    • 2002
  • Human endogenous retroviral long terminal repeats (LTRs) have been found to be coexpressed with sequences of genes located nearby. It has been suggested that the LTR elements have contributed to the structural change or genetic variation of human genome connected to various diseases. The HERV-W family has been identified in the cerebrospinal fluids and brains of individuals with schizophrenia. Using a cDNA library derived from a human brain, the HERV-W LTR elements were examined and five new LTR elements were identified. These elements were examined using a YAC clone panel from the Xq21.3 region linked to psychosis that was replicated on the Y chromosome after the separation of the chimpanzee and human lineages. Fourteen elements of the HERV-W LTR were identified in that region. Those LTR elements showed a high degree of sequence similarity ($91.8-99.5\%$) with previously reported HERV-W LTR. A phylogenetic tree obtained from the neighbor-joining method revealed that new HERV-W LTR elements were closely related to the AXt000960, AF072504, and AF072506 from the GenBank database. The data indicates that several copy numbers of the HERV-W LTR elements exist on the Xq21.3 region and are also expressed in the human brain. These LTR elements need to be further investigated as potential leads to neuropsychiatric diseases.

Use of the Non-electrophoretic Method to Detect Testis Specific Protein Gene for Sexing in Preimplantation Bovine Embryos

  • Huang, Jinming;You, Wei;Wu, Naike;Tan, Xiuwen
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권6호
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    • pp.866-871
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    • 2007
  • Testis-specific protein (TSPY) is a Y-specific gene, with up to 200 copy numbers in bulls. In order to make bovine embryo sexing under farm condition more feasible, the possibility of using a non-electrophoretic method to detect the TSPY gene for sexing bovine early embryos was examined. Primers were designed to amplify a portion of the TSPY gene and a common gene as an internal control primer. PCR optimization was carried out using a DNA template from bovine whole blood. Furthermore, embryo samples were diagnosed by this method and the sexing results were contrasted with those of the Loop-Mediated Isothermal Amplification (LAMP) method. The results showed that TSPY was as reliable a sexing method as LAMP. Forty-three morula and blastocyst embryos collected from superovulated donor dairy cattle were sexed by this method, and twenty-one embryos judged to be female embryos were transferred non-surgically to recipients 6 to 8 days after natural estrus. Out of 21 recipients, 9 were pregnant (42.86%) and all delivered female calves. The results showed that the sex predicted by this protocol was 100% accurate. In conclusion, the TSPY gene was a good male specific marker and indicated that a non-electrophoretic method was feasible and accurate to detect the TSPY gene for sexing preimplantation bovine embryos.

Tetrahydropteridines possess antioxidant roles to guard against glucose-induced oxidative stress in Dictyostelium discoideum

  • Park, Seon-Ok;Kim, Hye-Lim;Lee, Soo-Woong;Park, Young Shik
    • BMB Reports
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    • 제46권2호
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    • pp.86-91
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    • 2013
  • Glucose effects on the vegetative growth of Dictyostelium discoideum Ax2 were studied by examining oxidative stress and tetrahydropteridine synthesis in cells cultured with different concentrations (0.5X, 7.7 g $L^{-1}$; 1X, 15.4 g $L^{-1}$; 2X, 30.8 g $L^{-1}$) of glucose. The growth rate was optimal in 1X cells (cells grown in 1X glucose) but was impaired drastically in 2X cells, below the level of 0.5X cells. There were glucose-dependent increases in reactive oxygen species (ROS) levels and mitochondrial dysfunction in parallel with the mRNA copy numbers of the enzymes catalyzing tetrahydropteridine synthesis and regeneration. On the other hand, both the specific activities of the enzymes and tetrahydropteridine levels in 2X cells were lower than those in 1X cells, but were higher than those in 0.5X cells. Given the antioxidant function of tetrahydropteridines and both the beneficial and harmful effects of ROS, the results suggest glucose-induced oxidative stress in Dictyostelium, a process that might originate from aerobic glycolysis, as well as a protective role of tetrahydropteridines against this stress.

A Cyan Fluorescent Protein Gene (cfp)-Transgenic Marine Medaka Oryzias dancena with Potential Ornamental Applications

  • Vu, Nguyen Thanh;Cho, Young Sun;Lee, Sang Yoon;Kim, Dong Soo;Nam, Yoon Kwon
    • Fisheries and Aquatic Sciences
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    • 제17권4호
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    • pp.479-486
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    • 2014
  • To evaluate their potential utility as an ornamental organism, novel transgenic marine medaka Oryzias dancena strains with a highly vivid fluorescent phenotype were established through transgenesis of a cyan fluorescent protein gene (cfp) driven by the endogenous fast skeletal myosin light chain 2 gene (mlc2f) promoter. The transgenic marine medaka strains possessed multiple copies of transgene integrants and passed their fluorescent transgenes successfully to subsequent generations. Transgenic expression in skeletal muscles at both the mRNA and phenotypic levels was, overall, dependent upon transgene copy numbers. In the external phenotype, an authentic fluorescent color was dominant in the skeletal muscles of the transgenic fish and clearly visible to the unaided eye. The phenotypic fluorescent color presented differentially in response to different light-irradiation sources; the transgenics displayed a yellow-green color under normal daylight or white room light conditions, a strong green-glowing fluorescence under ultraviolet light, and a cyan-like fluorescence under blue light from a light-emitting diode.