• Proceedings of the Korean Society of Life Science Conference
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• 2001.02a
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• pp.84-84
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• 2001

• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1714-1727
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• 2015

This work describes a new strategy for optimal design of Multiplex-PCR primer sequences. The process is based on the Particle Swarm Optimization-Simplex algorithm (Mult-PSOS). Diverging from previous solutions centered on heuristic tools, the Mult-PSOS is selfconfigured because it does not require the definition of the algorithm's initial search parameters. The successful performance of this method was validated in vitro using Multiplex-PCR assays. For this validation, seven gene sequences of the most prevalent bacteria implicated in urinary tract infections were taken as DNA targets. The in vitro tests confirmed the good performance of the Mult-PSOS, with respect to infectious disease diagnosis, in the rapid and efficient selection of the optimal oligonucleotide sequences for Multiplex-PCRs. The predicted sequences allowed the adequate amplification of all amplicons in a single step (with the correct amount of DNA template and primers), reducing significantly the need for trial and error experiments. In addition, owing to its independence from the initial selection of the heuristic constants, the Mult-PSOS can be employed by non-expert users in computational techniques or in primer design problems.

• Journal of Plant Biotechnology
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• v.6 no.1
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• pp.25-37
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• 2004

Successful genetic transformation of plants requires non-chimeric selection of transformed tissues and their subsequent regeneration. With rare exceptions, most transformation protocols still rely heavily on antibiotics for selecting transgenic cells that contain an antibiotic-degrading selectable marker gene. Here, the morphogenic capacity of in-vitro explants of chrysanthemnum and tobacco stems and leaves (control and transgenic) changed with the addition of aminoglycoside antibiotics (AAs), In a test of 6 AAs, phytotoxicity occurred at concentrations of 10 to 25 and 50 to 100$\mu\textrm{g}$ $mL^{-1}$ in chrysanthemum and tobacco explants, respectively. Light conditions as well as explant source and size also had significant effects. The use of transverse thin cell layers (tTCLs), in conjunction with high initial AA selection levels, supported the greatest regeneration of transgenic material (adventitious shoots or callus) and the lowest number of escapes. Flow-cytometric analyses revealed no endodu-plication in chrysanthemum, even at high AA levels. However, this phenomenon was observed in tobacco calli(8C or more), even at low AA concentrations (i.e., 5 to 10 $\mu\textrm{g}$ mL$^{-1}$ ).

• Journal of Forest and Environmental Science
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• v.34 no.3
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• pp.224-234
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• 2018

An efficient method to select drought tolerant Korean native plants using in vitro culture system was established in this study. While the plant growths and root inductions of each plant were proportionately affected by concentrations of mannitol on in vitro culturing seven plant species to test tolerance to osmotic stress, growth index (GI) and number of root induction of Chrysanthemi zawadskii var. latilobum and Dianthus chinensis var. semperflorens plantlets were higher than the others in 125mM mannitol. In test with polyethylene glycol (PEG), plantlets of C. zawadskii var. latilobum and D. chinensis var. semperflorens showed higher GI and number of root induction than the others in 33.3mM. On testing whether the well grown plants under osmotic stress are tolerant to virtual drought stress, there were significant differences in the withering rates of C. zawadskii var. latilobum and D. chinensis and those of were Aster yomena and Centaurea cyanus after 12 days without watering. It was found that significantly lower stomata numbers were shown in both drought tolerant plants than the sensitive plants. Averages of the stomata circumferences and the stomata area in the plantlets of the tolerant species were larger than those of the sensitive plants D. chinensis var. semperflorens showed the lowest transpiration level per unit area. The highest stomatal area per unit area was found in C. zawadskii, followed by D. chinensis var. semperflorens, Aster yomena and C. cyanus. In conclusion, C. zawadskii var. latilobum and D. chinensis var. semperflorens were more tolerant to drought than other two species. Furthermore in vitro selection was successfully used to screen drought tolerance species of native plant species.

• Journal of Plant Biotechnology
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• v.5 no.1
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• pp.51-57
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• 2003

Doubled haploids have long been recognized as a valuable tool in plant breeding since it not only offers the quickest method of advancing heterozygous breeding lines to homozygosity, but also increased the selection efficiency over conventional procedures due to better discrimination between genotypes within any one generation. Salt tolerant mutants were obtained in rice the variety, 'Hawsungbyeo', through in vitro mutagenesis of in vitro cultured anther-derived calli. Various doses (30, 50, 70 and 90 Gy) of gamma ray were applied to investigate the effect of radiation on callus formation on medium containing 1% NaCl, green plant regeneration, frequency of selected doubled haploid mutants and of the salt tolerant screen. It was demonstrated that the dose of 30 and 50 Gy gamma rays had significant effects on callus formation, regeneration and selection of salt tolerance. No tolerant lines were obtained from non-mutagenized cultures. From gamma ray irradiated cultures, five tolerant lines ($M_2$generation) at germination stage and 13 tolerant lines ($M_3$genoration) at seedling stage were obtained. The frequency of salt tolerant mutants indicates that anther culture applied in connection with gamma rays is an effective way to improve salt tolerance.

• Bulletin of the Korean Chemical Society
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• v.26 no.12
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• pp.2033-2037
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• 2005

In vitro selection was used to isolate $Mg^{2+}$-dependent self-cleaving ribozymes with cis-cleavage activity from a pre-tRNA library having 40-mer random sequences attached to 5'-end of E. coli $tRNA^{Phe}$. After 8 rounds of SELEX (Systematic Evolution of Ligands by Exponential Enrichment), RNA molecules which can self-cleave at the high concentration of $Mg^{2+}$ were isolated. The selected ribozymes can carry out the self-cleavage reaction in the presence of 100 mM $Mg^{2+}$ but not in 10 mM $Mg^{2+}$. The cleavage sites of the ribozymes are located at +3 and +4 of $tRNA^{Phe}$, compared with +1 position of 5'-end cleavage site of pre-tRNA by RNase P. New RNA constructs deprived of its D stem-loop, anticodon stem-loop, variable loop and T stem-loop, respectively showed the cleavage specificity identical to a ribozyme having the intact tRNA structure. Also, the new ribozyme fused with both a ribozyme and $tRNA^{Leu}$ showed the cleavage activities at the various sites within its sequences, different from two sites of position +3 and +4 observed in the ribozyme with $tRNA^{Phe}$. Our results suggest that the selected ribozyme is not structural-specific for tRNA.

• Proceedings of the Korean Society of Life Science Conference
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• 2006.09a
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• pp.55.1-55.1
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• 2006

• Journal of Radiation Industry
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• v.6 no.3
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• pp.199-204
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• 2012

• Journal of Embryo Transfer
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• v.10 no.1
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• pp.33-44
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• 1995

Genetic changes from improving female's reproductive rate through in vitro fertilization of large number of oocytes were studied. The breeding scheme employed was multiple ovulation and embryo transfer of juveniles and adults. Both balanced and unbalanced matings were examined for the four closed progeny population sizes, 10$^3$, 10$^4$, 105, 106. In balanced matings, all selected sires and dams were mated to each other(cross-classified mating) while unbalanced matings allowed selected dams and sires mated partially, eg. unbalanced matings allowed averages of .5 and .25 progeny per each mating. Various numbers of selected sires and dams were also examined in both balanced and unbalanced matings. In all mating schemes, selection of males and females was restricted to he one from each fullsib family to reduce the rate of inbreeding. The model calculations were deterministic and accounted for the effects of selection and inbreeding on loss of the genetic variation in succeeding generations. Balanced rectangular mating schemes, where more donors were selected than sires, resulted in larger selection responses than balanced square mating schemes, where equal numbers of sires and donors were selected, and unbalanced rectangular mating. The first round selection responses from the balanced rectangular matings of juvenile MOET, eg. number of progeny per mating equals 2 with 10 sires selected, were 1.192, 1.406, 1.580 and 1.735 times larger than the first round selection responses from the balanced square mating schemes for the given four progeny population sizes, 10$^3$, 10$^4$, 105 and 106, respectively. Similar results were obtained in adult MOET breeding schemes. However, balanced square matings gave greater selection responses than the unbalanced rectangular matings.

• Journal of Embryo Transfer
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• v.23 no.2
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• pp.93-100
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• 2008

This study was conducted to establish an in vitro maturation (IVM) system by selection of efficient macromolecule in the porcine in vitro production (IVP) technology. To choose the efficient macromolecules in the development of porcine embryos, the effects of 3 kinds of macromolecules (porcine serum; PS, porcine follicular fluid; pFF, and polyvinyl alcohol; PVA) supplemented in IVM media on the maturation, cleavage, and development rates to blastocyst of parthenogenetic activation (PA) and in vitro fertilization (IVF) embryos were examined. The maturation rates of porcine oocytes in media supplemented with PS were significantly higher than those with pFF and PVA (92.4% vs. 85.4%, 77.1%; p<0.05). In the cleavage and development to blastocyst rates, supplement with PS or pFF in the IVM media was more effective than PA. However, there were no significant differences in cleavage and development to blastocyst between PS and pFF group. From the results of this study, it was demonstrated that PS was optimal macromolecule in the porcine IVM media.