• 제목/요약/키워드: functional gene

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남성과 여성에서 XIST 유전자의 후성학적 비교 연구 (Epigenetic Study of XIST Gene from Female and Male Cells by Pyrosequencing)

  • 김환희;윤여진;송민애;이수만
    • Clinical and Experimental Reproductive Medicine
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    • 제37권1호
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    • pp.25-31
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    • 2010
  • 목 적: X 염색체 불활성화는 여성과 남성 사이에 X 염색체의 유전자 발현 유지를 위해 여성의 X 염색체 중 하나가 불활성화 되는 현상이다. 이러한 X 염색체 불활성화는 해독되지 않는 XIST 유전자에 의해 조절된다. XIST 유전자는 오직 불활성화된 X 염색체 에서만 발현되고, 활성화된 X 염색체 에서는 발현되지 않는다. 따라서 체세포에서 활성화된 X 염색체의 XIST 유전자는 promoter 부분이 메틸화 되어있고, 불활성화된 X 염색체에서는 메틸화가 거의 되어 있지 않다. 연구방법: 본 연구에서는 정상 여성과 정상 남성의 XIST 유전자의 promoter와 5'-end 지역의 메틸화 차이를 측정하기 위해 정상여성과 남성의 혈액에서 DNA를 추출하여 파이로시퀀싱 (Pyrosequencing) 방법을 통해 XIST 유전자의 총 8부분의 CpG 영역 (-1696, -1679, -1475, -1473, -1469, +947, +956, +971)을 분석하였다. 결 과: 총 8부분의 CpG 영역을 분석한 결과, promoter 부분인 CpG 1-5 영역 (-1696, -1679, -1475, -1473, -1469)에서는 여성과 남성의 메틸화 정도에 차이가 없었다. 그러나 5'-end 부분인 CpG6-8 영역 (+947, +956, +971)에서는 여성이 45.2% 49.9% 44.2%, 남성이 90.6%, 96.7%, 87.8%으로 메틸화 정도가 차이를 나타냈다. 결 론: 따라서 본 연구에 사용한 방법은 XIST 유전자의 메틸화 패턴의 차이를 기존의 방법보다 신속하고 정확하게 분석할 수 있다는 장점이 있기 때문에 유용하게 사용될 수 있을 것이다.

Computational analysis of large-scale genome expression data

  • Zhang, Michael
    • 한국생물정보학회:학술대회논문집
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    • 한국생물정보시스템생물학회 2000년도 International Symposium on Bioinformatics
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    • pp.41-44
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    • 2000
  • With the advent of DNA microarray and "chip" technologies, gene expression in an organism can be monitored on a genomic scale, allowing the transcription levels of many genes to be measured simultaneously. Functional interpretation of massive expression data and linking such data to DNA sequences have become the new challenges to bioinformatics. I will us yeast cell cycle expression data analysis as an example to demonstrate how special database and computational methods may be used for extracting functional information, I will also briefly describe a novel clustering algorithm which has been applied to the cell cycle data.

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Molecular and Biochemical Studies on the DNA Replication of Bacteriophage T7: Functional Analysis of Amino-terminal Region of Gene 2.5 Protein

  • Kim, Young-Tae;Lee, Sung-Gu;Kim, Hak-Jun
    • BMB Reports
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    • 제28권6호
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    • pp.484-489
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    • 1995
  • The product of bacteriophage T7 gene 2.5 is a single-stranded DNA binding protein and plays an important role in T7 DNA replication, recombination, and repair. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth (Kim and Richardson, 1993). The C-terminal truncated gene 2.5 protein ($GP2.5-{\Delta}21C$) cannot substitute for wild-type gene 2.5 protein in vivo; suggesting that the C-terminal domain of gene 2.5 protein is essential for protein-protein interactions (Kim and Richardson, 1994; J. Biol. Chem. 269, 5070-5078). Truncated gene 2.5 proteins lacking 19 residues ($GP2.5-{\Delta}19N$) and 39 residues ($GP2.5-{\Delta}39N$) from the amino-terminal domain were constructed by in vitro mutagenesis. $GP2.5-{\Delta}19N$ can support the growth of T7 phage lacking gene 2.5 while $GP2.5-{\Delta}39N$ cannot substitute for wild-type gene 2.5 protein in vivo; however, its ability to bind to single-stranded DNA is not affected. These results clearly demonstrate that the 20~39 amino-terminal region of gene 2.5 protein is required for T7 growth in vivo but may not be involved in DNA binding activity.

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Assessment of Risks and Benefits of Using Antibiotics Resistance Genes in Mesenchymal Stem Cell-Based Ex-Vivo Therapy

  • Narayan Bashyal;Young Jun Lee;Jin-Hwa Jung;Min Gyeong Kim;Kwang-Wook Lee;Woo Sup Hwang;Sung-Soo Kim;Da-Young Chang;Haeyoung, Suh-Kim
    • International Journal of Stem Cells
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    • 제16권4호
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    • pp.438-447
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    • 2023
  • Recently, ex-vivo gene therapy has emerged as a promising approach to enhance the therapeutic potential of mesenchymal stem cells (MSCs) by introducing functional genes in vitro. Here, we explored the need of using selection markers to increase the gene delivery efficiency and evaluated the potential risks associated with their use in the manufacturing process. We used MSCs/CD that carry the cytosine deaminase gene (CD) as a therapeutic gene and a puromycin resistance gene (PuroR) as a selection marker. We evaluated the correlation between the therapeutic efficacy and the purity of therapeutic MSCs/CD by examining their anti-cancer effect on co-cultured U87/GFP cells. To simulate in vivo horizontal transfer of the PuroR gene in vivo, we generated a puromycin-resistant E. coli (E. coli/PuroR) by introducing the PuroR gene and assessed its responsiveness to various antibiotics. We found that the anti-cancer effect of MSCs/CD was directly proportional to their purity, suggesting the crucial role of the PuroR gene in eliminating impure unmodified MSCs and enhancing the purity of MSCs/CD during the manufacturing process. Additionally, we found that clinically available antibiotics were effective in inhibiting the growth of hypothetical microorganism, E. coli/PuroR. In summary, our study highlights the potential benefits of using the PuroR gene as a selection marker to enhance the purity and efficacy of therapeutic cells in MSC-based gene therapy. Furthermore, our study suggests that the potential risk of horizontal transfer of antibiotics resistance genes in vivo can be effectively managed by clinically available antibiotics.

A Comprehensive Review of Emerging Computational Methods for Gene Identification

  • Yu, Ning;Yu, Zeng;Li, Bing;Gu, Feng;Pan, Yi
    • Journal of Information Processing Systems
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    • 제12권1호
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    • pp.1-34
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    • 2016
  • Gene identification is at the center of genomic studies. Although the first phase of the Encyclopedia of DNA Elements (ENCODE) project has been claimed to be complete, the annotation of the functional elements is far from being so. Computational methods in gene identification continue to play important roles in this area and other relevant issues. So far, a lot of work has been performed on this area, and a plethora of computational methods and avenues have been developed. Many review papers have summarized these methods and other related work. However, most of them focus on the methodologies from a particular aspect or perspective. Different from these existing bodies of research, this paper aims to comprehensively summarize the mainstream computational methods in gene identification and tries to provide a short but concise technical reference for future studies. Moreover, this review sheds light on the emerging trends and cutting-edge techniques that are believed to be capable of leading the research on this field in the future.

Novel SNP in the coding region of the FTO gene is associated with marbling score in Hanwoo (Korean cattle)

  • Chung, Eui-Ryong
    • Journal of Animal Science and Technology
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    • 제56권8호
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    • pp.27.1-27.6
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    • 2014
  • The fat mass and obesity associated (FTO) gene plays an important role in the regulation of energy homeostasis, fat deposition and obesity. For this reason, the FTO gene is a physiological and functional candidate gene for carcass and meat quality traits in beef cattle. The objectives of this study were to identify SNPs in the exonic regions of FTO gene and to evaluate the association of these SNPs with carcass traits in Hanwoo (Korean cattle). In this study, we newly identified two exonic SNPs in Hanwoo population. The g.125550A > T SNP was located in exon 3 and the g.175675C > T SNP was located in exon 6. Genotyping of the two SNP markers was carried out using PCR-RFLP analysis in Hanwoo steers to evaluate their association with carcass traits. As a result, g.125550A > T SNP genotype was significantly associated with effects on marbling score. Animals with the AA and TT homozygous genotypes had a significantly higher marbling score (p < 0.001) than those with AT heterozygous genotype, and this was significant after Bonferroni correction of the significance threshold (p = 0.003). Dominance effect was also observed for the marbling score (P < 0.05) with higher marbling score of homozygous animals. However, no significant associations with meat quality traits were observed for the g.175675C > T SNP. Our results suggest that the exonic SNP g.125550A > T in the FTO gene may be used as a DNA marker for the selection of Hanwoo with higher marbling.

Baicalin을 처리한 HL-60 백혈병 세포주에서 대규모 유전자 분석 발현 연구 (Studies on Gene Expression of baicalin treated in HL-60 cell line using High-throughput Gene Expression Analysis Techniques)

  • 김봉주;차민호;전병훈;윤용갑;윤유식
    • 동의생리병리학회지
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    • 제18권5호
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    • pp.1291-1300
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    • 2004
  • Baicalin, a biologically active flavonoid form the roots of Scutallaria baicalensis (Skullcap), have been reported to not only function as anti-oxidants but also cause anticancer effect. We investigated the mechanism of baicalin-induced cytotoxicity and the macro scale gene expression analysis in leukemia cell line, HL-60 cells. Baicalin (10 μM) were used to treat the cells for 6h, 12h, 24h, 48h and 72h. In a human cDNAchip study of 65,000 genes evaluated 6, 12, 24, 48. 72 hours after treated with Baicalin in HL-60 cells. Hierarchical cluster against the genes which showed expression changes by more than two fold. One hundred one genes were grouped into 6 clusters according to their profile of expression by a hierarchical clustering algorithm. For genes differentially expressed in response to baicalin treatment, we tested functional classes based on Gene Ontology (GO) terms. This study provides the most comprehensive available survey of gene expression changes in response to baicalin treatment in HL-60 cell line.

Gene Expression Profiling in Rice Infected with Rice Blast Fungus using SAGE

  • Kim, Sang-Gon;Kim, Sun-Tae;Kim, Sung-Kun;Kang, Kyu-Young
    • The Plant Pathology Journal
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    • 제24권4호
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    • pp.384-391
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    • 2008
  • Rice blast disease, caused by the pathogenic fungus Magnaporthe grisea, is a serious issue in rice (Oryza sativa L.) growing regions of the world. Transcript profiling in rice inoculated with the fungus has been investigated using the transcriptomics technology, serial analysis of gene expression (SAGE). Short sequence tags containing sufficient information which are ten base-pairs representing the unique transcripts were identified by SAGE technology. We identified a total of 910 tag sequences via the GenBank database, and the resulting genes were shown to be up-regulated in all functional categories under the fungal biotic stress. Compared to the compatible interaction, the stress and defense genes in the incompatible interaction appear to be more up-regulated. Particularly, thaumatin-like gene (TLP) was investigated in determining the gene and protein expression level utilizing Northern and Western blotting analyses, resulting in an increase in both the gene and the protein expression level which arose earlier in the incompatible interaction than in the compatible interaction.