• 제목/요약/키워드: Copy number

검색결과 464건 처리시간 0.021초

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

  • 홍억기
    • KSBB Journal
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    • 제9권3호
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    • pp.339-345
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    • 1994
  • 포도당 억제현상은 유전자 조작 및 induder에 의해 감소될 수 있다. UA8G와 GALl TATA box 사 이에서의 유전자 삭제는 포도당 억제현상을 줄이고 갈락토스가 존재하지 않는 조건에서 지속적인 유전자 발현을 도모했다. 상대적 inducer의 양(갈락토스/포 도당 농도의 비)은 유전자 발현 및 포도당 억제현상 에 영향을 주었다. 포도당 억제현상은 상대적 inducer의 양이 증가함에 따라 2-5배 정도 감소하였다. 또한 유전자 발현은 플라즈미드의 수에 좌우된다. 배지에 갈락토스만 았을 경우 유전자 발현은 플라즈 마드의 수가 증가함에 따라 증가하였다. 반면에 배지에 포도당과 갈락토스가 함께 있는 경우 (2% G Glu+2% Gal), 플라즈미드의 수는 유전자 발현에 별다른 영향을 주지 못했다. 그러나 높은 상대적 m­d ducer 양이 배지에 있는 경우 (0.4% Glu+0.8% Gal), 플라즈미드의 수가 증가함에 따라 유전자 발 현이 증가하였다. 즉, 포도당 억제현상을 줄임으로써 유전자 발현효율을 높이고자 할 때 갈락토스의 농도 를 증가시키는 경우보다는 포도당의 농도를 낮춤으 로써 상대적 inducer의 양음 높여 유전자 발현을 유 도하는 방법이 보다 효율적인 것으로 나타났다.

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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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    • 제65권4호
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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
    • 대한의생명과학회지
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    • 제15권3호
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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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    • 제16권1호
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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.

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

  • Chung, Kung-Sook;Kim, Choon-Kwang;Kim, Kyu-Won
    • 미생물학회지
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    • 제31권1호
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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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    • 제8권3호
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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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    • 제52권6호
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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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    • 제17권6호
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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.

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

  • 김승민;정준범
    • 한국어병학회지
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    • 제31권2호
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    • pp.101-107
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    • 2018
  • 여윔증상이 나타난 양식장(farm-B 및 farm-C)의 넙치 및 여윔증상이 나타나지 않은 양식장(farm-A) 넙치의 각 내부 장기(신장, 장, 비장, 뇌 및 간)를 대상으로 점액포자충 Parvicapsula sp.의 양적 분석을 real-time PCR 방법을 사용하여 각각 실시하였다. 여윔증상을 보였던 farm-C의 넙치 신장에서 가장 높은 DNA copy number ($1.7{\times}10^7copies/mg$ tissue)를 보였고, farm-B의 넙치에서는 모든 내부 장기에서 낮은 수치가 나타났으며, farm-A의 넙치에서는 모든 내부 장기에서 음성 결과를 나타내었다. 동일한 시료를 사용한 PCR 및 병리조직학적 분석에서도 real-time PCR에서의 결과와 같은 양상을 보였다.

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

  • 백정훈;임제탁
    • 전자공학회논문지S
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    • 제35S권10호
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    • pp.52-63
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    • 1998
  • 본 논문에서는 기존의 멀티캐스트 패킷 교환기의 문제점에 대한 개선 방안을 제안한다. 오버플로우에 의해 야기 되는 입력포트의 공평성 문제는 임의의 입력 포트로부터 복사 수를 가산 할 수 있는 동적 시작점 결정기에 의해 해결된다. 시작점은 매 타임 슬롯 마다 입력 버퍼의 점유도와 이전 타임 슬롯의 오버플로우를 기반으로 가변 된다. 입력 버퍼의 점유도를 이용함으로서 제안된 복사망은 기존의 방식에 비해 입력 트래픽의 변동에 대하여 우수한 적응성을 제공한다. 동적 시작점 결정기는 입력 트래픽의 양에 따라 복사 요청의 수를 제어하며 이것은 복사망의 전체 스루풋을 제고하는 필수적인 기능 이다. 오버플로우 발생시에 멀티캐스트 스위치의 스루풋을 향상시키는 호-분할(call-splitting) 방식도 동적 시작점 결정기에 의해 제공된다. 동적 시작점 결정기의 하드웨어는 고속 동작에 적합한 단순한 구조로 도출된다. 제안된 방식의 성능 평가를 위해 다양한 트래픽에 대한 모의 실험 결과가 제공된다.

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