• Title/Summary/Keyword: copy number

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Effects of Glucose Repression and Plasmid Copy Number on Cloned Gene Expression in Recombinant Yeast (재조합 효모에서의 포도당 억제와 Plasmid 수가 유전자 발현에 미치는 영향)

  • 홍억기
    • KSBB Journal
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    • v.9 no.3
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    • pp.339-345
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    • 1994
  • Deletions between UASG and the GALI TATA box reduced glucose repression and allowed constitutive expression of the gene product in the absence of galactose. The relative inducer level (ratio of galactose/glucose concentrations) affected the extent of gene expression and glucose repression. Glucose repression was reduced by a factor of 2 to 5 as the relative inducer level increased. In the medium containing galactose only, induction of ${\beta}$-galactosidase synthesis by galactose increased with plasmid copy number. On the contrary, plasmid copy number did not affect significantly ${\beta}$-galactosidase synthesis in the medium containing both glucose and galactose (2% glucose+2% galactose), which might be due to glucose repression caused by high glucose concentration. However, when the medium contained the relatively high inducer level (0.4% glucose+0.8% galactose), ${\beta}$-galactosidase synthesis increased with plasmid copy number, indicating that the beneficial effect of higher galactose concentration was weaker than the repressive effect of higher glucose concentration.

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Quantitative analysis of mitochondrial DNA in porcine-mouse cloned embryos

  • Hyeonyeong Shin;Soyeon Kim;Myungyoun Kim;Jaeeun Lee;Dongil Jin
    • Journal of Animal Science and Technology
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    • v.65 no.4
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    • pp.767-778
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    • 2023
  • The aim of the research is to identify that porcine oocytes can function as recipients for interspecies cloning and have the ability to develop to blastocysts. Furthermore each mitochondrial DNA (mtDNA) in interspecises cloned embryos was analyzed. For the study, mouse-porcine and porcine-porcine cloned embryos were produced with mouse fetal fibroblasts (MFF) and porcine fetal fibroblasts (PFF), respectively, introduced as donor cells into enucleated porcine oocytes. The developmental rate and cell numbers of blastocysts between intraspecies porcine-porcine and interspecies mouse-porcine cloned embryos were compared and real-time polymerase chain reaction (PCR) was performed for the estimate of mouse and porcine mtDNA copy number in mouse-porcine cloned embryos at different stages.There was no significant difference in the developmental rate or total blastocyst number between mouse-porcine cloned embryos and porcine-porcine cloned embryos (11.1 ± 0.9%, 25 ± 3.5 vs. 10.1 ± 1.2%, 24 ± 6.3). In mouse-porcine reconstructed embryos, the copy numbers of mouse somatic cell-derived mtDNA decreased between the 1-cell and blastocyst stages, whereas the copy number of porcine oocyte-derived mtDNA significantly increased during this period, as assessed by real-time PCR analysis. In our real-time PCR analysis, we improved the standard curve construction-based method to analyze the level of mtDNA between mouse donor cells and porcine oocytes using the copy number of mouse beta-actin DNA as a standard. Our findings suggest that mouse-porcine cloned embryos have the ability to develop to blastocysts in vitro and exhibit mitochondrial heteroplasmy from the 1-cell to blastocyst stages and the mouse-derived mitochondria can be gradually replaced with those of the porcine oocyte in the early developmental stages of mouse-porcine cloned embryos.

Evaluation of DNA Extraction Methods from Low Copy Number (LCN) DNA Samples for Forensic DNA Typing

  • Eom, Yong-Bin
    • Biomedical Science Letters
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    • v.15 no.3
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    • pp.229-232
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    • 2009
  • DNA isolation for PCR-based short tandem repeat (STR) analysis is essential to recover high yields of amplifiable DNA from low copy number (LCN) DNA samples. There are different methods developed for DNA extraction from the small bloodstain and gloves, commonly found at crime scenes. In order to obtain STR profiles from LCN DNA samples, DNA extraction protocols, namely the automated $iPrep^{TM}$ $ChargeSwitch^{(R)}$ method, the automated $QIAcube^{TM}$ method, the automated $Maxwell^{(R)}$ 16 DNA $IQ^{TM}$ Resin method, and the manual $QIAamp^{(R)}$ DNA Micro Kit method, were evaluated. Extracted DNA was quantified by the $Quantifiler^{TM}$ Human DNA Quantification Kit and DNA profiled by $AmpFISTR^{(R)}$ $Identifiler^{(R)}$ Kit. Results were compared based on the amount of DNA obtained and the completeness of the STR profiles produced. The automated $iPrep^{TM}$ $ChargeSwitch^{(R)}$ and $QIAcube^{TM}$ methoas produced reproducible DNA of sufficient quantity and quality trom the dried blood spot. This two automated methods showed a quantity and quality comparable to those of the forensic manual standard protocols normally used in our laboratory. In our hands, the automated DNA extraction method is another obvious choice when the forensic case sample available is bloodstain. The findings of this study indicate that the manual simple modified $QIAamp^{(R)}$ DNA Micro Kit method is best method to recover high yields of amplifiable DNA from the numerous potential sources of LCN DNA samples.

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Is Mitochondrial DNA Copy Number Associated with Clinical Characteristics and Prognosis in Gastric Cancer?

  • Lee, Hyunsu;Lee, Jae-Ho;Kim, Dong-Choon;Hwang, IlSeon;Kang, Yu-Na;Gwon, Gi-Jeong;Choi, In-Jang;Kim, Shin
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.1
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    • pp.87-90
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    • 2015
  • Alterations in mitochondrial DNA (mtDNA) have been studied in various cancers. However, the clinical value of mtDNA copy number (mtCN) alterations in gastric cancer (GC) is poorly understood. In the present study, we investigated whether alterations in mtCNs might be associated with clinicopathological parameters in GC cases. mtCN was measured in 109 patients with GC by quantitative real-time PCR. Then, correlations with clinicopathological characteristics were analyzed. mtCN was elevated in 64.2% of GC tissues compared with paired, adjacent, non-cancerous tissue. However, the observed alterations in mtCN were not associated with any clinicopathological characteristics, including age, gender, TN stage, Lauren classification, lymph node metastasis, and depth of invasion. Moreover, Kaplan-Meier survival curves revealed that mtCN was not significantly associated with the survival of GC patients. In this study, we demonstrated that mtCN was not a significant marker for predicting clinical characteristics or prognosis in GC.

Stability of Recombinant Plasmids Carrying the stb Locus of E. coli IncFII NR1 Plasmid in E. coli and Yeast (대장균과 효모에서 Escherichia coli IncFII NR1 플라스미드의 stb 좌위를 포함하는 재조합 플라스미드의 안정성에 관한 연구)

  • Chung, Kung-Sook;Kim, Choon-Kwang;Kim, Kyu-Won
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.37-43
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    • 1993
  • The effect of stb locus of E. COLI IncFII plasmid NR1 on the stability of chimeric plasmids was investigated. First, we have isolated the stability locus (stb) from E. coli NR1 plasmid and then inserted into the three different vectors, pUC8, YRp17 and YEp24. By examining their stability in E. coli and yeast, we showed that the recombinant plasmids containing stb locus were resonably stable. Also, by comparing the amounts of the rDNA fragments per haploid genome with those of the plasmid fragments, we showed they copy number of recombinant plasmids was not increased. Consequently, the stb locus of E. coli IncFII plasmid NR1 stabilized the chimeric plasmids but did not affect the replication or copy number of plasmids.

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A Genome-wide Association Study of Copy Number Variation in Hematological Parameters in the Korean Population

  • Kim, Ka-Kyung;Cho, Yoon-Shin;Cho, Nam-H.;Shin, Chol;Kim, Jong-Won
    • Genomics & Informatics
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    • v.8 no.3
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    • pp.122-130
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    • 2010
  • Abnormal hematological values are associated with various disorders including cancer and cardiovascular, metabolic, infectious, and immune diseases. We report the copy number variations (CNVs) in clinically relevant hematological parameters, including hemoglobin level, red and white blood cell counts, platelet counts, and red blood cell (RBC) volume. We describe CNVs in several loci associated with these hematological parameters in 8,842 samples from Korean population-based studies. The data that we evaluated included four RBC parameters, one platelet parameter, and one associated with total white blood cell (WBC) count, exceeding the genome-wide significance. We show that CNVs in hematological parameters are associated with some loci, different from previously associated loci reported in single nucleotide polymorphism (SNP) association studies.

Large-Scale Copy-Number Alterations in Chicken Ovarian Cancer

  • Seo, Hee-Won;Choi, Jin-Won;Yun, Tae-Won;Lee, Hong-Jo;Kim, Hee-Seung;Song, Yong-Sang;Song, Gwon-Hwa;Han, Jae-Yong
    • Journal of Animal Science and Technology
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    • v.52 no.6
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    • pp.491-498
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    • 2010
  • Copy-number variation (CNV) in particular genomic segments owing to deletions or duplications can induce changes in cellular gene expression patterns and may increase susceptibility to diseases such as cancer. The aim of this study was to examine CNVs related to the incidence of epithelial ovarian cancer in chickens. Genomic DNA was extracted from blood cells and cancerous ovaries collected from four 120-week-old White Leghorn chickens and were used for array-based comparative genome hybridization (CGH) analysis. As a result, 25 amplified and 10 deleted CNV regions were detected in chicken ovarian cancer. Of these, 10 amplified and two deleted CNV regions contained genes associated with human ovarian cancer. Our study using a chicken model may provide a better understanding of human epithelial ovarian cancer.

Genetic Abnormalities in Oral Leukoplakia and Oral Cancer Progression

  • Kil, Tae Jun;Kim, Hyun Sil;Kim, Hyung Jun;Nam, Woong;Cha, In-Ho
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.6
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    • pp.3001-3006
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    • 2016
  • Background: The cancer progression of oral leukoplakia is an important watchpoint in the follow-up observation of the patients. However, potential malignancies of oral leukoplakia cannot be estimated by histopathologic assessment alone. We evaluated genetic abnormalities at the level of copy number variation (CNV) to investigate the risk for developing cancer in oral leukoplakias. Materials and Methods: The current study used 27 oral leukoplakias with histological evidence of dysplasia. The first group (progressing dysplasia) consisted of 7 oral lesions from patients with later progression to cancer at the same site. The other group (non-progressing dysplasia) consisted of 20 lesions from patients with no occurrence of oral cancer and longitudinal follow up (>7 years). We extracted DNA from Formalin-Fixed Paraffin-Embedded (FFPE) samples and examined chromosomal loci and frequencies of CNVs using Taqman copy number assays. Results: CNV frequently occurred at 3p, 9p, and 13q loci in progressing dysplasia. Our results also indicate that CNV at multiple loci-in contrast to single locus occurrences-is characteristic of progressing dysplasia. Conclusions: This study suggests that genetic abnormalities of the true precancer demonstrate the progression risk which cannot be delineated by current histopathologic diagnosis.

Quantitative analysis of a myxosporean parasite, Parvicapsula sp. detected from emaciated olive flounder, Paralichthys olivaceus in Korea (국내 여윔 넙치에서 검출된 점액포자충 Parvicapsula sp.의 정량적 분석)

  • Kim, Seung Min;Jeong, Joon Bum
    • Journal of fish pathology
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    • v.31 no.2
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    • pp.101-107
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    • 2018
  • Quantitative analysis of a myxosporean parasite, Parvicapsula sp. in internal organs (kidney, intestine, spleen, brain and liver) from non-emaciated (farm-A) or emaciated (farm-B and farm-C) olive flounder Paralichthys olivaceus were performed by real-time PCR. The highest DNA copy number ($1.7{\times}10^7copies/mg$ tissue) was detected in kidney of the emaciated olive flounder from farm-C, while the DNA copy number was below detection limit in all the organs of the olive flounder from farm-B. There was not positive result in all of organs from olive flounder in farm-A. PCR and histopathological analysis were also performed using the same specimen and showed same results as those by real-time PCR.

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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