• Title/Summary/Keyword: 유전자 이동

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Expression of Sodium/iodide Symporter Transgene in Neural Stem Cells (신경줄기세포(HB1.F3)에서 나트륨옥소 공동수송체 도입유전자 발현)

  • Kim, Yun-Hui;Lee, Dong-Soo;Kang, Joo-Hyun;Lee, Yong-Jin;Chung, June-Key;Lee, Myung-Chul
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.1
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    • pp.99-108
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    • 2004
  • Purpose: The ability to noninvasively track the migration of neural progenitor cells would have significant clinical and research implications. We generated stably transfected F3 human neural progenitor cells with human sodium/iodide symporter (hNIS) for noninvasively tracking F3. In this study, the expression patterns of hNIS gene in F3-NIS were examined according to the cultured time and the epigenetic modulation. Materials and Methods: F3 human neural stem cells had been obtained from Dr. Seung U. Kim (Ajou University, Suwon, Korea). hNIS and hygromycin resistance gene were linked with IRES (Internal Ribosome Entry Site) under control of CMV promoter. This construct was transfected to F3 with Liposome. To investigate the restoration of hNIS gene expression in F3-NIS, cells were treated with demethylating agent (5-Azacytidine) and Histone deacetylase inhibitor (Trichostatin A: TSA). The expression of hNIS was measured by I-125 uptake assay and RT-PCR analysis. Results: The iodide uptake of the F3-NIS was higher 12.86 times than F3 cell line. According to the cell passage number, hNIS expression in F3-NIS gradually diminished. After treatment of 5-Azacytidine and TSA with serial doses (up to $20{\mu}M$, up to 62.5nM, respectively) for 24 hours, I-125 uptake and mRNA of hNIS in F3-NIS were increased. Conclusion: These results suggest that hNIS transfected F3 might undergo a change in its biological characters by cell passage. Therefore, the gene ex[ressopm of exogenous gene transferred human stem cell might be affected to the epigenetic modulation such as promoter methylation and Histone deacetylation and to the cell culture conditions.

Up-regulation of Pluripotency-related Genes in Human Amniotic Fluid-derived Stem Cells by ESRRB Conjugated with Cell-Penetrating Peptide (인간 양수 유래 줄기세포에서 세포투과단백질을 이용한 ESRRB 단백질의 직접도입에 의한 전분화능 관련 유전자의 발현 조절)

  • Jo, Jung-Hyun;Lee, Yu-Sun;Oh, Mi-Hee;Ko, Jung-Jae;Cheon, Yong-Pil;Lee, Dong-Ryul
    • Development and Reproduction
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    • v.14 no.4
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    • pp.243-251
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    • 2010
  • ESRRB (Estrogen related receptor $\beta$) is an orphan receptor, and have a role on maintaining the undifferentiated state and self-renewal of pluripotent stem cell as a transcription factor which regulates the expression of OCT4 and NANOG genes. Also, Feng et al. (2009) reported that Esrrb, Oct4 and Sox2 could induce pluripotent stem cell from somatic cells. The aim of the present study was to develop the direct delivery system of human ESRRB protein into human amniotic fluid-derived stem cells (AFSCs) and to analyze the effect of ESRRB on the regulation of pluripotency-related genes. Human ESRRB has three isoforms arisen by alternative splicing. We cloned short-form ESRRB and made a fusion protein of ESRRB and R7 for an efficient protein transfer to cell. R7 as cell-penetrating peptide(CPP) can help to transfer ESRRB into cells. R7-ESRRB-His6 protein was observed in the cytoplasm and nuclei within 5 hours after treatment. Also, we could observe R7-ESRRB-His6 protein only in the nuclei within 24 hours. Realtime PCR showed that ESRRB increased expression of OCT4 and NANOG as well as SOX2 gene. Therefore, we demonstrated that R7-ESRRB-His6 proteins were efficiently transferred into the nuclei of AFSCs and work well as a possible transcription factor.

Molecular Cloning of the Gene Coding for 3-Isopropylmalate Dehydrogenase of Kluyveromyces fragilis (Kluyveromyces fragilis의 LEU gene의 Cloning)

  • 박성희;이동선;우주형;김종국;홍순덕
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.305-308
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    • 1990
  • In order to clone the gene coding for 3-isopropylmalate dehydrogenase of Muyveromyces fragilis, a shuttle plasmid vector pHNll4 was used. It can serve as a cloning vector in Saccharomyces cerevisiae DBY746 for other Sau3AI-cleaved DNA segment of Kluyveromyces fragilis. Two cloned fragments which complement the leu2 mutation of Saccharomyces cerevisiae and E, coli were obtained. Their length was 4.4 kb an 3.5 kb, and their orientation was opposite each other. From the fact that the two recombinant plasmids were expressed in Saccharomyces cerevisiae and E, coli, probably the two inserts had the promoter of Ktuyveromyces fi-agilis and that of Kluyveromyces fiagilis was efficiently assosiated with RNA polymerase of Saccharomyces cerevisiae and E. coli. According to the result of Southern hybridization, we thought that the cloned fragment has low homology with 3-isopropylmalate dehydrogenase coding region of E. coli and Saccharomyces cerevisiae.

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Cloning and Sequence Analysis of the Kinesin Gene in Schizosaccharomyces pombe (Schizosaccharomyces pombe의 Kinesin 유전자의 클로닝과 염기서열분석)

  • 정재욱;최성민;김형배;이명석
    • Korean Journal of Microbiology
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    • v.35 no.1
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    • pp.18-24
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    • 1999
  • Kinesin has been discovered in Saccharomyces cerevisiae, Aspergillus nidulans, and Drosophila melanogaster and it has major roles in the movemenl of chromosomes and separation of spindle poles. In this study, a gene encoding kinesin heavy chain in Schizosaccharo~n)~ces pombe was cloned by using the polymerase chain reaction with degenerated primcrs corresponding to highly conserved regions of the kinesin heavy chain motor domain. The kinesin gene in S pombe contains an open reading frame of 2496 base pairs and encodes a kinesin prolein of 832 amino acids with a molecular weight of 96 kd. From thc comparison of the predictcd amino acids of the newly cloned kinesin, the kinesin in S. pornbe belongs to the C-terminal motor subfamily of kincsin-related protein.

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Genetic Algorithm-Based Watermarking in Discrete Wavelet Transform Domain (유전자 알고리듬을 사용한 웨이블릿 기반 워터마킹)

  • Lee Dong-Eun;Kim Tae-Kyung;Lee Seong-Won;Paik Joon-Ki
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.43 no.4 s.310
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    • pp.108-115
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    • 2006
  • This paper presents a watermarking algorithm in the discrete wavelet transform domain using evolutionary algorithm. The proposed algorithm consists of wavelet-domain watermark insertion and genetic algorithm-based watermark extraction. More specifically watermark is inserted to the low-frequency region of wavelet transform domain, and watermark extraction is efficiently performed by using the evolutionary algorithm. The proposed watermarking algorithm is robust against various attacks such as JPEG and JPEG2000 image compression and geometric transformations.

Generic Scheduling Method for Distributed Parallel Systems (분산병렬 시스템에서 유전자 알고리즘을 이용한 스케쥴링 방법)

  • Kim, Hwa-Sung
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.28 no.1B
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    • pp.27-32
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    • 2003
  • This paper presents the Genetic Algorithm based Task Scheduling (GATS) method for the scheduling of programs with diverse embedded parallelism types in Distributed Parallel Systems, which consist of a set of loosely coupled parallel and vector machines connected via high speed networks The distributed parallel processing tries to solve computationally intensive problems that have several types of parallelism, on a suite of high performance and parallel machines in a manner that best utilizes the capabilities of each machine. When scheduling in distributed parallel systems, the matching of the parallelism characteristics between tasks and parallel machines rather than load balancing should be carefully handled with the minimization of communication cost in order to obtain more speedup. This paper proposes the based initialization methods for an initial population and the knowledge-based mutation methods to accommodate the parallelism type matching in genetic algorithms.

A Study on Incremental Evolution of Neural Network based on Cellular Automata (셀룰라 오토마타 기반 신경망의 점증적 진화에 관한 연구)

  • 송금범;조성배
    • Proceedings of the Korean Information Science Society Conference
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    • 1998.10c
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    • pp.348-350
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    • 1998
  • 시뮬레이션 환경이나 실제 환경에서 이동 로봇의 제어에 관한 많은 연구가 진행되어 왔다. 이러한 연구 중에서 이동 로봇이 장애물을 피한다거나, 움직이는 물체를 잡는 등의 행동을 유전자 알고리즘 등의 진화 알고리즘으로 만들어내는 연구가 최근 활발하다. 이전의 연구에서는 셀룰라 오토마타 상에서 진화의 방법으로 신경망을 성장시키는 모델을 제시하고, 그 유용성을 입증하고자 이동로봇의 제어에 적용하여 나름대로 만족할 만한 결과를 얻을 수 있었다. 그러나 이러한 진화의 방법은 환경에 제한된 제어기를 만들어 내는 문제점이 있어 본 논문에서는 점증적인 진화의 방법을 이용하여 좀더 다양한 환경에 적응할 수 있는 제어기를 만들어 내고자 한다. 점증적 방법은 초기에 간단한 행동으로 해결할 수 있는 환경에 맞도록 제어기를 진화시킨 다음, 점차 복잡한 행동이 요구되는 환경에서 제어기를 점증적으로 진화시킨다. 실험 결과, 점증적 진화의 방법이 좀더 효율적으로 로봇을 진화시키고 환경의 변화에 보다 강한 것을 알 수 있었다.

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Cell Migratory Induction by Expression of Angiogenin and Vascular Endothelial Growth Factor in Resveratrol Treated HeLa Cells (Resveratrol 처리한 HeLa세포에서 angiogenin과 vascular endothelial growth factor 발현유도에 따른 세포이동촉진)

  • Joe, I-Seul;Jeong, Sin-Gu;Cho, Goang-Won
    • Journal of Life Science
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    • v.24 no.4
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    • pp.337-342
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    • 2014
  • Resveratrol (RSV), a natural polyphenolic compound, is a modulator for cell division and cell migration, and has diverse beneficial properties. Angiogenin (ANG) and vascular endothelial growth factor (VEGF) are considered to be important mechanisms for cell proliferation, angiogenesis, the formation of tubular structures, and migration. In this study, we investigated whether RSV has a migratory effect in HeLa cells. When cells were treated with $0{\sim}50{\mu}M$ of RSV for 24 hr, the expression of ANG and VEGF was significantly increased in a dose dependent manner measured by real-time PCR. Similarly, we performed time dependent experiments for $50{\mu}M$ RSV treated cells and identified the optimized time at 24 hr. The increased expression in RSV treated cells was confirmed by Western blot analysis. To examine the toxic effects of RSV at the determined conditions, MTT assays were performed. The viabilities were unchanged for $0{\sim}50{\mu}M$ RSV treated cells, while they decreased at $100{\mu}M$ RSV. To examine the effect of migration in RSV treated cells, we performed a wound-healing assay. The migratory rates were significantly enhanced in the RSV treated group. In this study, we found that RSV induces an increase in the expression of migration factors ANG, VEGF, and enhances cell migration for the determined conditions.

Reidentification of Comamonas sp. Strain DJ-12 and Analysis of its pcbABC2D2 Genes Responsible for Degradation of 4-Chlorobiphenyl. (Comamonas sp. Strain DJ-12 의 재동정 및 4-Chlorobiphenyl 분해유전자 pcbABC2D2 의 분석)

  • 이준훈;박동우;강철희;채종찬;이동훈;김치경
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.121-126
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    • 2004
  • Comamonas sp. strain DJ-12 is a 4-chlobiphenyl(4CB)-degrading bacterium that was reidentified from Pseudomonas sp. DJ-12. The genomic DNA was isolated from the strain DJ-12 and amplified by PCR with primers for cloning pcbABCD genes responsible for degradation of 4CB. The amino acid sequences deduced from the nucleotide sequences of pcbA1, pcbA2, pcbA3, pcbA4, pcbB, pcbC2, and pcbD2 genes showed 91, 87, 99, 87, 97, 90 and 87% homologies with those of Pseudomonas sp. KKS102, respectively. The pcbC1D1 genes that are involved in the degradation of (4-chloro)1,2-dihydroxybiphenyl produced from 4CB by pcbAB gene products were previously reported in the recombinant plasmid pCU1 from Pseudomonas sp. DJ-12. However, the pcbC2D2 genes in the plasmid pCT4 and pCT5 cloned from Comamonas sp. DJ-12 in this study showed 51 and 62% homologies with those of pcbC1D1 in their nucleotide sequences. The pcbC1D1 and pcbC2D2 genes were found by Southern hybridization to be located at different loci on the chromosome of DJ-12 strain. These results indicate that Comamonas sp. strain DJ-12 has two different sets of pcbCD genes responsible for deg-radation of (4-chloro)1,2-dihydroxybiphenyl.

Conservative Genes of Less Orthologous Prokaryotes (Orthologs 수가 적은 원핵생물들의 보존적 유전자)

  • Lee, Dong-Geun
    • Journal of Life Science
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    • v.27 no.6
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    • pp.694-701
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    • 2017
  • Mycoplasma genitalium represents the smallest genome among mono-cultivable prokaryotes. To discover and compare the orthologs (conservative genes) among M. genitalium and 14 prokaryotes that are uncultivable and have less orthologs than M. genitalium, COG (clusters of orthologous groups of protein) analyses were applied. The analyzed prokaryotes were M. genitalium, one hyperthermophilic exosymbiotic archaeon Nanoarchaeum equitans, four intracellular plant pathogenic eubacteria of Candidatus Phytoplasma genus, and nine endosymbiotic eubacteria of phloem- and xylem-feeding insects. Among 367 orthologs of M. genitalium, 284 orthologs were conservative between M. genitalium and at least one other prokaryote. All 15 prokaryotes commonly have 29 orthologs, representing the significance of proteins in life. They belong to 25 translation-related, including 22 ribosomal proteins, 3 subunits of RNA polymerase, and 1 protein-folding-related. Among the 15 prokaryotes, 40 orthologs were only found in all four Candidatus Phytoplasma. The other nine Candidatus, all endosymbionts with insects, showed only a single common COG0539 (ribosomal protein S1), representing the diversity of orthologs among them. These results might provide clues to understand conservative genes in uncultivable prokaryotes, and may be helpful in industrial areas, such as handling prokaryotes producing amino acids and antibiotics, and as precursors of organic synthesis.