• Title/Summary/Keyword: ITS copy number

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A Penalized Spline Based Method for Detecting the DNA Copy Number Alteration in an Array-CGH Experiment

  • Kim, Byung-Soo;Kim, Sang-Cheol
    • The Korean Journal of Applied Statistics
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    • v.22 no.1
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    • pp.115-127
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    • 2009
  • The purpose of statistical analyses of array-CGH experiment data is to divide the whole genome into regions of equal copy number, to quantify the copy number in each region and finally to evaluate its significance of being different from two. Several statistical procedures have been proposed which include the circular binary segmentation, and a Gaussian based local regression for detecting break points (GLAD) by estimating a piecewise constant function. We propose in this note a penalized spline regression and its simultaneous confidence band(SCB) approach to evaluate the statistical significance of regions of genetic gain/loss. The region of which the simultaneous confidence band stays above 0 or below 0 can be considered as a region of genetic gain or loss. We compare the performance of the SCB procedure with GLAD and hidden Markov model approaches through a simulation study in which the data were generated from AR(1) and AR(2) models to reflect spatial dependence of the array-CGH data in addition to the independence model. We found that the SCB method is more sensitive in detecting the low level copy number alterations.

Genome Architecture and Its Roles in Human Copy Number Variation

  • Chen, Lu;Zhou, Weichen;Zhang, Ling;Zhang, Feng
    • Genomics & Informatics
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    • v.12 no.4
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    • pp.136-144
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    • 2014
  • Besides single-nucleotide variants in the human genome, large-scale genomic variants, such as copy number variations (CNVs), are being increasingly discovered as a genetic source of human diversity and the pathogenic factors of diseases. Recent experimental findings have shed light on the links between different genome architectures and CNV mutagenesis. In this review, we summarize various genomic features and discuss their contributions to CNV formation. Genomic repeats, including both low-copy and high-copy repeats, play important roles in CNV instability, which was initially known as DNA recombination events. Furthermore, it has been found that human genomic repeats can also induce DNA replication errors and consequently result in CNV mutations. Some recent studies showed that DNA replication timing, which reflects the high-order information of genomic organization, is involved in human CNV mutations. Our review highlights that genome architecture, from DNA sequence to high-order genomic organization, is an important molecular factor in CNV mutagenesis and human genomic instability.

An advanced tool, droplet digital PCR (ddPCR), for absolute quantification of the red-tide dinoflagellate, Cochlodinium polykrikoides Margalef (Dinophyceae)

  • Lee, Hyun-Gwan;Kim, Hye Mi;Min, Juhee;Kim, Keunyong;Park, Myung Gil;Jeong, Hae Jin;Kim, Kwang Young
    • ALGAE
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    • v.32 no.3
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    • pp.189-197
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    • 2017
  • To quantify the abundance of the harmful dinoflagellate Cochlodinium polykrikoides in natural seawaters, we developed the innovative procedure using a droplet digital PCR (ddPCR) with C. polykrikoides-specific primers targeting the internal transcription sequence (ITS). The abundance of C. polykrikoides was estimated by the specific copy number of target ITS DNA segments per cell in cultures and natural water samples. The copy number per C. polykrikoides cell as acquired by ddPCR was $157{\pm}16$, which was evaluated against known cell numbers through a simplified protocol preparing DNAs. The abundances of C. polykrikoides in the waters of different locations estimated by ddPCR agreed with the number of cells visually counted under a microscope. This protocol was used to measure the abundance of C. polykrikoides close to and further off the southern coast of Korea in August of 2016 and 2017. The practical application showed that this method can reduce time for analysis and increase accuracy.

재조합 효모를 이용한 항혈전 단백질 히루딘 발효 생산공정의 최적화

  • Kim, Myeong-Dong;Gang, Hyeon-A;Lee, Sang-Gi;Seo, Jin-Ho
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.99-102
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    • 2001
  • Recombinant Saccharomyces cerevisiae strains harboring various copy numbers of hirudin gene were developed to study dependency of hirudin expression level on its gene copy number. A linear relationship between the copy number of hirudin expression cassette and hirudin expression level was observed up to 10 copies. A double <5-integration vector truncated wi 디 1 the unnecessary bacterial genes before yeast transformation showed a four-fold increase in transformation efficiency and a 1.3-fold enhancement in hirudin expression level compared with a single <5 system. Gratuitous hirudin expression strain was developed by disrupting the GALl gene of S. cerevisiae. Glucose that was fed in a limited manner effectively supported cell growth and hi겨din expression by the gratuitous strain. Effects of methanol concentrations on hirudin production in recombinant Hansenula polymorpha were investigated in continuous and fed-batch cultures. At a steady-state of continuous culture, an optimum methanol concentration of 1.7 g/L was determined at a dilution rate of 0.18 $h^{-1}$ with 1.8 mg/L ${\cdot}$ h hirudin productivity.

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Quantification of the ichthyotoxic raphidophyte Chattonella marina complex by applying a droplet digital PCR

  • Juhee, Min;Kwang Young, Kim
    • ALGAE
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    • v.37 no.4
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    • pp.281-291
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    • 2022
  • Quantifying the abundance of Chattonella species is necessary to effectively manage the threats from ichthyotoxic raphidophytes, which can cause large-scale mortality of aquacultured fish in temperate waters. The identification and cell counting of Chattonella species have been conducted primarily on living cells without fixation by light microscopy because routine fixatives do not retain their morphological features. Species belonging to the Chattonella marina complex, including C. marina and C. marina var. ovata, had high genetic similarities and the lack of clear morphological delimitations between the species. To estimate the abundance of C. marina complex in marine plankton samples, we developed a protocol based on the droplet digital polymerase chain reaction (ddPCR) assay, with C. marina complex-specific primers targeting the internal transcribed spacer (ITS) region of the rDNA. Cell abundance of the C. marina complex can be determined using the ITS copy number per cell, ranging from 25 ± 1 for C. marina to 112 ± 7 for C. marina var. ovata. There were no significant differences in ITS copies estimated by the ddPCR assay between environmental DNA samples from various localities spiked with the same number of cells of culture strains. This approach can be employed to improve the monitoring efficiency of various marine protists and to support the implementation of management for harmful algal blooms, which are difficult to analyze using microscopy alone.

Cloning, Base Sequence Determination and Homology Analysis of Replication Controlling cop Gene of R-plasmid pSBK203 Isolated from Staphylococcus aureus DHI (Staphylococcus aureus DH1에서 분리된 R-plasmid pSBK203의 복제조절 유전자 cop의 Cloning, 염기서열 결정 및 상동성 분석)

  • Park, Seung-Moon;Byeon, Woo-Hyeon
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.115-119
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    • 1994
  • Replication control region of pSBK203, a chloramphenicol acetyltransferase conferring plasmid from Staphylococus aureus was cloned and its nucleotide sequence has been determined. Base sequence homology of this copy control region with those of plasmids belonging to pT181 family was obtained and analyzed. Copy number of four copy mutants derived by addtion or deletion of nucleotides in unique XbaI recognition site in copy control region of pSBK203 was also determined.

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The copy networks controlling the copy number according to the fluctuations of the input traffics for an ATM Multicast Switch (입력 트래픽의 특성에 따라 복사 수가 제어되는 ATM 멀티캐스트 스위치 복사 망)

  • Paik, Jung-Hoon;Lim, Chae-Tak
    • Journal of the Korean Institute of Telematics and Electronics S
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    • v.35S no.10
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    • pp.52-63
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    • 1998
  • In this paper, several improvements to a copy network proposed previously for multicast packet switching are described. The improvements provide a solution to some problems inherent in multicasting. The input fairness problem caused by overf low is solved by a dynamic starting point decider(DSD), which can calculate running sums of copy requests starting from any input port. The starting point is changed adaptively in every time slot based on both the fill level of the input buffers in current time slot and the overflow situations of the previous time slot. Using the fill level of the conventional network. The DSD also provides the function of regulating overall copy requests according to the amount of input traffics. This is an essential function in improving overall throughputs of the copy networks. The throughput of a multicast switch can be improved substantially if partial service of copy request is implemented when overflow occurs. Call-splitting can also be implemented by the DSD in a straightforward manner. The hardware for the DSD is derived with the objective of simple architectures for the high speed operation. Simulation study of the copy network under various traffic conditions is presented to evaluate its performance.

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High-Resolution Microarrays for Mapping Promoter Binding sites and Copy Number Variation in the Human Genome

  • Albert Thomas
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2006.02a
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    • pp.125-126
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    • 2006
  • NimbleGen has developed strategies to use its high-density oligonucleotide microarray platform (385,000 probes per array) to map both promoter binding sites and copy number variation at very high-resolution in the human genome. Here we describe a genome-wide map of active promoters determined by experimentally locating the sites of transcription imitation complex binding throughout the human genome using microarrays combined with chromatin immunoprecipitation. This map defines 10,567 active promoters corresponding to 6,763 known genes and at least 1,196 un-annotated transcriptional units. Microarray-based comparative genomic hybridisation (CGH) is animportant research tool for investigating chromosomal aberrations frequently associated with complex diseases such as cancer, neuropsychiatric disorders, and congenital developmental disorders. NimbleGen array CGH is an ultra-high resolution (0.5-50 Kb) oligo array platform that can be used to detect amplifications and deletions and map the associated breakpoints on the whole-genome level or with custom fine-tiling arrays. For whole-genome array CGH, probes are tiled through genic and intergenic regions with a median probe spacing of 6 Kb, which provides a comprehensive, unbiased analysis of the genome.

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Quantitative Analysis of Bacillus amyloliquefaciens GR4-5 in Soil (Bacillus amyloliquefaciens GR4-5 균주의 토양 내 정량 분석)

  • Kim, Dayeon;Kim, Byung-Yong;Ahn, Jae-Hyung;Weon, Hang-Yeon;Kim, Sung-Il;Kim, Wan-Gyu;Song, Jaekyeong
    • Korean Journal of Organic Agriculture
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    • v.23 no.4
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    • pp.847-858
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    • 2015
  • Bacillus amyloliquefaciens GR4-5 was isolated from the rhizosphere soil of Korean ginseng and displayed broad-spectrum suppression of ginseng root rot pathogens. The survivability of B. amyloliquefaciens GR4-5 in soil was investigated under three different conditions; indoor, outdoor - of which soil was put in 14 mL tube after treatment - and field environments. Soil samples were collected over a four-week period from three experimental designs, and assessed for 16S rRNA gene copy number by quantitative polymerase chain reaction (qPCR). In outdoor condition, the 16S rRNA gene copy number of Bacillus spp. was 8.35 log copies g $soil^{-1}$ immediately after the GR4-5 treatment. Two weeks later, the 16S rRNA gene copy number of Bacillus spp. (6.70 log copies g $soil^{-1}$) was similar to that of the control (6.38 log copies g $soil^{-1}$). In indoor condition, the 16S rRNA gene copy number of Bacillus spp. maintained in a certain level for a longer period than those in outdoor and field. The 16S rRNA gene copy number of Bacillus spp. in field experiment was reduced faster than that of outdoor condition. Our results show that B. amyloliquefaciens GR4-5 can survive in bulk soil for 1 week, indicating its potential use as a biocontrol agent following 7 day application intervals. This study presents that outdoor microcosm system design could be a useful method to assess easily the survivability of beneficial microorganisms.

Write Request Handling for Static Wear Leveling in Flash Memory (SSD) Controller

  • Choo, Chang;Gajipara, Pooja;Moon, Il-Young
    • Journal of information and communication convergence engineering
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    • v.12 no.3
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    • pp.181-185
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    • 2014
  • The lifetime of a solid-state drive (SSD) is limited because of the number of program and erase cycles allowed on its NAND flash blocks. Data cannot be overwritten in an SSD, leading to an out-of-place update every time the data are modified. This result in two copies of the data: the original copy and a modified copy. This phenomenon is known as write amplification and adversely affects the endurance of the memory. In this study, we address the issue of reducing wear leveling through efficient handling of write requests. This results in even wearing of all the blocks, thereby increasing the endurance period. The focus of our work is to logically divert the write requests, which are concentrated to limited blocks, to the less-worn blocks and then measure the maximum number of write requests that the memory can handle. A memory without the proposed algorithm wears out prematurely as compared to that with the algorithm. The main feature of the proposed algorithm is to delay out-of-place updates till the threshold is reached, which results in a low overhead. Further, the algorithm increases endurance by a factor of the threshold level multiplied by the number of blocks in the memory.