• Journal of Life Science
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• v.20 no.5
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• pp.789-793
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• 2010

Artificial chromosome manipulation and amplification of single-copy yeast artificial chromosome (YAC) are usually required in order to use YACs for applications such as physical mapping and functional analysis in eukaryotes. We designed and implemented a Simultaneous YAC Manipulation-Amplification (SYMA) system that combines the copy number amplification system of YAC with a convenient YAC manipulation system. To achieve the desired split and to amplify a YAC clone-harboring plant chromosome, a pBGTK plasmid containing a conditional centromere and thymidine kinase (TK) gene was constructed as a template to amplify the splitting fragment via PCR. By splitting, new 490-kb and 100-kb split YACs containing the elements for copy number amplification were simultaneously generated from a 590-kb YAC clone. The 100-kb split YAC was then successfully amplified 14.4-fold by adding 3 mg/ml sulfanilamide and $50\;{\mu}g/ml$ methotrexate (S3/M50) as inducing substances.

• Microbiology and Biotechnology Letters
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• v.50 no.3
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• pp.436-440
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• 2022

In this study, yeast artificial chromosome Insert (YAC) harboring genes which related xylan metabolism was constructed by using chromosome manipulation technique. For efficient chromosome manipulation, each splitting fragment (DNA module) required for splitting process was prepared and these DNA modules were transformed into Saccharomyces cerevisiae strain YKY164. By two-rounds chromosome splitting, yeast chromosome VII (1,124 kb) was split 887 kb-YAC, 45 kb-mini YAC and 198 kb-YAC and YKY183 strain containing 18 chromosomes was constructed. Splitting efficiency for chromosome manipulation was 50- 78% and expression level of foreign genes on 45 kb-mini YAC and enzyme activity were indistinguishable from that of the YKY164 strain. Furthermore, xylan-degraded products by recombinant enzymes were confirmed and mini-yeast artificial chromosome maintained stable mitotic stability without chromosome loss during 160 generations.

• Journal of Photoscience
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• v.12 no.2
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• pp.75-82
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• 2005

Rumex acetosa L. is a dioecious flowering plant with well developed sex chromosome system: 2n = 12 + XX in the female plants and 2n = 12 + XY1Y2 in the male plants. To isolate sex-linked DNA, we carried out chromosome micromanipulation, followed by DOP-PCR, AFLP of the PCR products, reverse Southern hybridization and sequence analysis. From 500 AFLP specific clones, 13 X-chromosome and 5 Y-chromosome specific clones were obtained. Except one clone RADAX-239 ($\underline{R}umex\;\underline{a}-\underline{D}OP-PCR-\underline{A}FLP-\underline{Y}-chromosome\;specific$), all clones appear to be R. acetosa plant-specific sequences and non-coding sequences. Southern blot analysis using these clones could not discriminate genomic DNAs either from male or female plants. Results of this study imply that both autosome-origin and degeneration of sex chromosomes are prevalent in plant systems.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2001.10a
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• pp.25-25
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• 2001

Generation of transgenic fish acquiring the ability to express desirable phenotypes offers new possibilities for addressing fundamental biological questions, and can also attribute to enhanced aquaculture productivity. I describe here the recent research progress in my laboratory with particular emphasis on the development of fast-growing autotransgenic fish and its chromosome-set manipulation using our experimental organism, the mud loach, Misgurnus mizolepis. (omitted)

• Journal of Life Science
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• v.26 no.6
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• pp.646-650
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• 2016

The influence of chromosome number on cell growth and cell aging was investigated in various yeast strains that have many artificial chromosomes constructed using a chromosome manipulation technique. Host strain FY833 and the YKY18, YKY18R, YKY24, and YKY30 strains harboring 16 natural chromosomes, 18 chromosomes, 18 chromosomes containing rDNA chromosome, 24 chromosomes, and 30 chromosomes, respectively, were used, and the specific growth rate of each strain was compared. The specific growth rates in the YKY18 and YKY24 strains were indistinguishable from that in the host strain, while those of the YKY18R and YKY30 strains were reduced to approximately 25% and 40% of the host strain level, respectively. Subsequently, the replicative life span was examined to investigate the relationship between the number of chromosomes and cell aging, and the life span was decreased to approximately 14% and 45% of the host strain level in the YKY24 and YKY30 strains, respectively. Moreover, telomere length, well known as a senescence factor, was shorter and more diversified in the strain, showing decreased life span. Therefore, these results suggest the possibility that an increase in the number of chromosomes containing artificial chromosomes caused cell aging, and we expected these observations would be applied to improve industrial strain harboring of versatile and special artificial chromosomes.

• 제어로봇시스템학회:학술대회논문집
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• 2003.10a
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• pp.2435-2440
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• 2003

Manipulation of the nano/micro scale object has been a key technology in biology as the sizes of DNA, chromosome, nucleus, cell and embryo are within such order. For instance, for embryo cell manipulation, the cell injection is performed manually. The operator often spends over a year to carry out a cell manipulation project. Since the typical success rate of such operation is extremely low, automation of such biological cell manipulation has been asked. As the operator spends most of his time in finding the position of cell in the Petri dish and in injecting bio-material to the cell from the best orientation. In this paper, we propose a new strategy and a vision system, by which one can find, recognize and track nucleus, polar body, and zona pellucida of the embryo cell for automatic biomanipulation. The deformable template matching algorithm has been used in recognizing the nucleus and polar body of each cell. Result suggests that it outperforms the conventional methods.

• Korean Journal of Ichthyology
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• v.13 no.1
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• pp.85-88
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• 2001

Temperature-related cleavage rates and mitotic intervals (${\tau}_o$) in the far eastern catfish Silurus asotus were studied by averaging the duration of the second and third embryonic divisions to establish effective procedures for chromosome manipulations. At higher temperatures eggs developed faster and underwent more asynchronous development. Mitotic intervals for far eastern catfish were $21.5{\pm}1.4$ min at $22^{\circ}C$, $18.5{\pm}1.2$ min at $24^{\circ}C$, and $14.0{\pm}2.1$ min at $26^{\circ}C$. There was a strong negative correlation between ${\tau}_o$ and water temperature (Y=-1.85X+21.9, $R^2=0.9868$, where Y is ${\tau}_o$ and X is temperature).

• Development and Reproduction
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• v.16 no.4
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• pp.321-327
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• 2012

Korean bullhead (Pseudobagrus fulvidraco) was collected from the Kum River areas of Kangkyung-eup, Nonsan city, Chungcheongnam-do, Korea, from April to June, 2012 and was fertilized in order to observe egg development and temperature-related cleavage rates and mitotic intervals (${\tau}_0$). The fertilized eggs were separative, demersal and light yellowish with $1.5{\pm}0.06mm$ in diameter, and did not contain oil globules. The first cleavage stages were 90 min, 80 min, 60 min and 50 min at $21^{\circ}C$, $24^{\circ}C$, $27^{\circ}C$ and $30^{\circ}C$, respectively. At higher temperatures, eggs developed faster and underwent further identical development. For Korean bullhead, ${\tau}_0$ were $33.4{\pm}2.08$ min at $21^{\circ}C$, $31.5{\pm}3.06$ min at $24^{\circ}C$, $28.1{\pm}2.11$ min at $27^{\circ}C$ and $26.4{\pm}3.35$ min at $30^{\circ}C$. There were strong negative correlations between the $\tau_0$ and water temperatures at all points studied (Y=-1.13X+58.15, $R^2$=0.98, n=30, where Y is ${\tau}_0$ and X is temperature). The results obtained in this work will be helpful for chromosome manipulation by use of cleavage frequency data and ${\tau}_0$ data in Korean bullhead.

• Korean Journal of Microbiology
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• v.35 no.1
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• pp.28-34
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• 1999

Fragmentation vectors are used to analyze function and genomic structure of a gene of interest by creating deletion derivatives of large fragments of genomic DNA cloned as yeast artificial chromosomes (YACs). Herein, we developed a new hicistronic fragmentation vector that contains internal ribosomal entry sile (IRES) of encephalomyocarditis vin~s (EMCV) and $\beta$-galactosidase as a reporter gene. This vector system provides a novcl loo1 to analyze expression patterns of a gene of interest due to simultaneous expression of a target gene as well as $\beta$-galactosidase driven from a single message. In addition, the bicistronic fragmentation vector contains four rare-cutting restriction enzyme sites in the polycloning sites which can be used to conveniently insert any kinds of genes and therefore facilitates targeting DNA scgments into YAC by means of homologous recombination. This approach establishes a paradigm for manipulation of mammalian DNA segments and characterization of expression and regulatory regions of mammalian gene cloned as YAC.

• Korean Journal of Poultry Science
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• v.16 no.1
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• pp.1-8
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• 1989

This experiment was conducted to improve the performance of chickens by the precise separation and analysis of chromosomes which are integrated genetic materials, and by the use of gene manipulation techniques. Following are the main results obtained. 1. When the chromosomes were separated through the leucocyte culture and analyzed by Giemsa banding techniques (especially by the method in which 20 layers of banding patterns could be found in chromosome ＃1), the normal Patterns of chromosomes ＃l-9 and sex chromosomes, and the location of constitutive heterochromatin without any gene activities in all chromosomes were discovered. 2. To utilize the primodial germ cells (PGC) as the genetic vector which is one of the most important gene manipulation techniques, PGC's from triploid were transplanted to normal host embryos. Since the donor PGC's(3n) were found in the gonads of growing host embryos gene manipulation in poultry using PGC's, seemed to be possible.