• Development and Reproduction
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• v.4 no.2
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• pp.153-160
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• 2000

In this study, genetic variation in the 5 strains of 2N-cont, meiotic-G2N, mitotic-G2N and two types of clones with different genetic backgrounds was investigated by developing their tolerance to water temperature and salinity, which is a physiological trait, into a quantitative trait. The temperature was set at 19$^{\circ}C$, 22.5$^{\circ}C$, 25$^{\circ}C$ and 30$^{\circ}C$, each of which was combined with 0$\textperthousand$, 15$\textperthousand$ and 30$\textperthousand$ of salinity respectively, making 12 groups in all. In the mean survival time (MST), samples with 15$\textperthousand$ of salinity showed the longest survival time at all temperatures. The 2N-cont had the longest 126.16 h followed by clone-11 and clone-15 surviving for 113.22 h and 91.05 h respectively. Gynogenetic diploids showed the shortest 87,32 h and 36.56 h. At 22.5 and 25$^{\circ}C$, MST of each strain was significantly short, showing similar results to those of the groups at 19$^{\circ}C$. The 2N-cont had the longest MST while clones had a longer MST than gynogenetic diploids. This could be due to gynogenesis which causes homozygosis among malignant harmful genes, leading to its appearance in populations and resulting in early death in individuals with such genes. On the other hand, MST of clones was longer than that of gynogenetic fish. This could be because the 1st gynogenetic generation, which is a parental population, has already had its malignant genes removed, while the clones of the 2nd gynogenetic generation have had their superior genes fixed as well as their tolerance and survival improved. When temperature was raised to 22.5$^{\circ}C$ and 25$^{\circ}C$, increase in variation was observed in gynogenetic diploids and decrease in clones in 15$\textperthousand$ of salinity. This shows that such a trait is genetic to a certain extent. Consequently, if this character is developed into a quantitative trait and applied to selective breeding, it could be a useful character to secure superior strains and individuals, and also it would be possible to improve populations genetically through selection.

• Korean Journal Plant Pathology
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• v.12 no.4
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• pp.389-394
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• 1996

도열병균, Magnaporthe grisea, 균주간의 유전적 유연관계를 분석하고 그들의 유전에 관한 '기본 정보를 얻고자 DNA polymorphism 분석을 실시하였다. 기주가 다른 도열병 균주들이 공시되었고 cloning에 의해 벼 도열병균 KJ201레이스 균주로부터 생성된 임의 선발 genomic clone들이 공시균주들간의 polymorphism을 밝히기 위해 사용되었던 바 그중 repectitive sequence를 보유한 repeated copy clone 하나가 선발되었다. Clone pMJ6에 의해 밝혀진 repetitive sequence는 Southern hybridization시 벼 분리균주에는 약 30개, 다른 기주 분리균에도 20∼33개의 밴드를 형성하였다. 반면 피 분리균주에는 단지 두 개의 밴드만을 나타내 분리기주가 다른 균주간에 뚜렷한 polymorphism이 존재하였으며 parsimony 분석에서도 역시 아주 먼 cluster를 형성하여 피 분리균은 다른 기주 분리균과 유전적으로 상당히 먼 것으로 추정되었다. 공시균의 genomic DNA를 HindIII로 처리했을 때 pMJ6에 의한 밴드양상은 공시균을 EcoRI으로 처리했을 때의 MGR probe의 밴드 양상과 유사하여 이 repeated copy clone이 도열병균주간의 유전적 유연관계를 분석하는데 MGR 못지않게 유용할 것으로 보인다.

• Animal Systematics, Evolution and Diversity
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• v.11 no.2
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• pp.291-299
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• 1995

Sampels of Korean field mice (apodemus peninsulae peninsulae Thomas ) from six localities in Korea were used for the analyses of mitochondrial DNa (Mt DNA) fragment patterns resulted from the digestion with eight restriction enzymes. A total of 29 fragments were recognized and seven mtDNA clones were revealed. The nucleotide-sequence divergences (p) among the seven mtDNA clones ranged from 0.42% to 2.01%. Moreover, the seven clones were grouped into three major subgroups with the mean divergence value of 1.52% among them. One subgroup was composed of three clones of 18 sample from three localities (16, Cheongu: 1, Mt. Sobaek : 1, Mt. weolak) L another subgroup, three clones of eight samples from four localities (2, Cheongju ; 2 , Mt. Weolak ; 2, Mt. Gaya ; 2, haenam) ; and the last subgroup, one clone of two samples from Cheongju. Three subgroups were also distinct with one another in their mtDNA genotypes of Stu I and the former two subgroups differed from the last subgroup in their genotypes with Pvu II. Further analyses with additional samples from various localities in Korea appeared to be necessary in order to clarify the taxonomic status of the distinct mtDNA subgroups.

• Journal of Ecology and Environment
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• v.33 no.3
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• pp.245-251
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• 2010

We evaluated the interaction between Cd and Zn in the bioaccumulation of seven clones of Salix caprea, which were exposed both to Cd and Zn alone and to a combination of Cd and Zn. Cadmium (Cd) and Zn concentration in the four treatments were administered in the following order: root > leaf > stem, and obvious differences were noted among the treatments and clones. The leaf Cd concentration of clone BH2 and stem Cd concentration of clone BH5 in the combined Cd and Zn treatment group was increased by 62% and 110%, respectively, relative of that of the Cd alone treatment group. On the other hand, the leaf and stem Zn concentration of clone BH8 in the combined Cd and Zn treatment group was reduced by 66% and 61%, respectively, relative to that of the Zn alone treatment group. Translocation of Cd and Zn from the root was higher in the leaf than in the stem, and the combined Cd and Zn treatment stimulated the translocation of Cd from the root to the leaf and stem, whereas it suppressed the translocation of Zn from the root to the leaf and stem. Therefore, the interaction effects were considered strongly synergistic with Cd in the presence of Zn, but proved antagonistic to Zn in the presence of Cd in the combined Cd and Zn treatment group. The phytoremediation potentials of the seven clones, which were estimated from standard indices of Cd and Zn concentration in Cd and Zn alone and the combined Cd and Zn treatment groups, were highest in clone BH3, and lowest in clone BH5. Therefore, we recognize S. caprea as an appropriate material for phytoremediation, and this is particularly the case with clone BH3. However, further research will be required to evaluate the effects of Cd and Zn on the physiological changes as well as tolerance mechanisms against metal toxicity in S. caprea clones.

• Journal of Microbiology and Biotechnology
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• v.21 no.4
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• pp.333-340
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• 2011

Herbicide-tolerant Zoysia grass has been previously developed through Agrobacterium-mediated transformation. We investigated the effects of genetically modified (GM) Zoysia grass and the associated herbicide application on bacterial community structure by using culture-independent approaches. To assess the possible horizontal gene transfer (HGT) of transgenic DNA to soil microorganisms, total soil DNAs were amplified by PCR with two primer sets for the bar and hpt genes, which were introduced into the GM Zoysia grass by a callus-type transformation. The transgenic genes were not detected from the total genomic DNAs extracted from 1.5 g of each rhizosphere soils of GM and non-GM Zoysia grasses. The structures and diversities of the bacterial communities in rhizosphere soils of GM and non-GM Zoysia grasses were investigated by constructing 16S rDNA clone libraries. Classifier, provided in the RDP II, assigned 100 clones in the 16S rRNA gene sequences library into 11 bacterial phyla. The most abundant phyla in both clone libraries were Acidobacteria and Proteobacteria. The bacterial diversity of the GM clone library was lower than that of the non- GM library. The former contained four phyla, whereas the latter had seven phyla. Phylogenetic trees were constructed to confirm these results. Phylogenetic analyses of the two clone libraries revealed considerable difference from each other. The significance of difference between clone libraries was examined with LIBSHUFF statistics. LIBSHUFF analysis revealed that the two clone libraries differed significantly (P<0.025), suggesting alterations in the composition of the microbial community associated with GM Zoysia grass.

• Reproductive and Developmental Biology
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• v.31 no.4
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• pp.227-233
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• 2007

This study was conducted to examine the mRNA expression of apoptosis-related and imprinted genes and methylation pattern of the differentially methylated region (DMR) of H19 gene in day 35 of SCNT pig fetuses. The day 35 of natural mating (control) or cloned (clone) pig fetuses were recovered from uterus. Endometrium from dam and liver from fetus were obtained, respectively. mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. The Bcl-2 mRNA expression in clone was significantly lower than that of control (p<0.05). The mRNA expression of H19 gene in both endometrium and liver was significantly higher in clone than that of control, respectively (p<0.05). The level of IGF-2 mRNA in liver of clone was significantly lower than that of control (p<0.05), whereas the mRNA expression of IGF2-R gene in liver of clone was significantly higher than that of control (p<0.05). The DMR of H19 was lower methylation pattern in clone than that of control. These results suggest that the aberrant mRNA expression of apoptosis-related and imprinted genes and the lower DMR methylation pattern of imprinted gene may be closely related to the inadequate fetal development of cloned fetus.

• Journal of the Korean Society of Environmental Restoration Technology
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• v.10 no.5
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• pp.40-50
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• 2007

One-year-old rooted cuttings of ten poplar clones and one willow clone were planted in a riparian area in Osan. Survival rate, growth performance, biomass, vitality, defoliation, leaf damages by diseases and/or insects and stem borer damage of the poplar and willow clones have been investigated for three growing seasons. Average survival rate of all eleven clones was declined from 80.7% for the first year to 60.7% for the third year. At three years after planting, poplar clones Dorskamp, ST-148 and Eco-28 showed the best survival rate of 80%. For height and DBH growth, the poplar clone Ay-48 and the willow clone 131-25 were the highest 8.3m and 9.5cm, respectively. However, poplar clones 72-30 and 72-31 were lower than those of the other clones. Clones Ay-48 and 131-25 seemed to have strong vitality when compare to the other clones. No serious damages by diseases and insects were found in most clones. Clones Ay-48 and ST-148 were the most tolerant to various diseases and insects. Clone Ay-48 produced the largest biomass for individual and annual total biomass, 22.5kg and 18.7ton $ha^{-1}$, respectively. Clone Dorskamp showed the best adaptability, which was estimated with survival rate, biomass and damages by various diseases and insects in the riparian area and followed by clones Ay-48, 97-19 and Eco-28. As a consequence, the four clones seemed to be the best candidate poplar clones for the establishment of riparian woody buffer.

• Journal of Biosystems Engineering
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• v.26 no.4
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• pp.391-398
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• 2001

A robot system for clone replication and gridding, which is a preliminary state of the genome research, was developed and evaluated its performance. This gridding robot system consisted of 1) a gridding heat that replicated the clone, 2) a manipulator, as a part of body of robot, which transferred the gridding head along x-, y-, z-axis, 3) a well plate arranging board, 4) a sterilization unit, and 5) a control unit. Performance of the system was evaluated with 1) repeatability of the robot system, 2) clone replication efficiency, 3) time requirement of the replication, and 4) sterilization efficiency. The repeatability error of the robot system showed 0.219 mm and 0.094 mm in the direction of x- and y-axis, respectively. The success rate of the clone replication with the gridding head was 100% on the membrane filter. The time required for the replication was four minutes and fifty-five seconds from the four 96 well plates to a 384 well plate meanwhile the required time with well experienced hand labor was three minutes thirty-five seconds. The gridding operation of clone could not be done by hand labor and the required time with robot system for the gridding on the membrance filter with the control program 5$\times$5: 1 copy and 384 gridding pins was twenty minutes and twenty-five seconds. The efficiency of the sterilization was considered to be satisfactory since no growth of fungi was found around the area of replication in the membrane filter.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.237-243
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• 2008

Chemical and microbial characteristics of bacterial populations were investigated in a quercus and pine humus forest soil. Soil pH was $5.3\pm0.4$ and $4.1\pm0.9$ from each sample of a quercus and pine humus forest soil; C/N ratio of humus forest soil was $17.84\pm4.6%$ and $21.76\pm8%$, respectively. Total organic acid was investigated as 69.57 mM/g dry soil and 53.72 mM/g dry soil in each humus forest soil. Glutamine, pyruvate, succinate, lactic acid and acetic acid of pine humus forest soil were $1.5\sim4.5$ times higher than those of quercus humus forest soil. As we evaluated phylogenetic characteristics of bacterial populations by 16S rRNA-ARDRA analysis with DNA extracted from each humus forest soil. Based on the 16S rRNA sequences, 44 clone from ARDRA groups of quercus humus forest soil were classified into 7 phyla: ${\alpha},{\beta},{\gamma},{\delta}$-Proteobacteria, Acidobacteria, Actinobacteria, and Firmicutes. Thirty-two clone from ARDRA groups of pine humus forest soil were classified into 8 phyla: ${\alpha},{\beta},{\gamma}$-Proteobacteria, Acidobacteria, Bacteroides, Verrucomicrobia, Planctomycetes, and Gemmatomonadetes. According to PCA (Principal Component Analysis) based on 16S rRNA base sequence, there were three main groups of bacteria. All clone of Cluster I were originated from quercus humus forest soil, while 67% clone of Cluster II and 63% clone of Clusters III were separated from pine humus forest soil.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.2
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• pp.173-181
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• 2004

To delay the onset of apoptosis in the culture, transformed Tn 5B1-4 cells harboring anti-apoptotic genes, bcl-2 and baculovirus p35, have been established and analyzed for their anti-apoptotic ability in suspension culture using spinner flasks. In the suspension culture at agitation speeds of 100 rpm and 200 rpm, the cell growth of cell clone expressing Bcl-2 protein was much higher than other two clones and the maximum cell density of the clone was 6.0 ${\times}$ 10$^{6}$ cells/ml and 6.2 ${\times}$ 10$^{6}$ cells/ml at day three of the incubation. On the other hand, the cell growth of cell clone expressing baculovirus protein P35 was much higher than other two clones in suspension culture at agitation speed of 300 rpm and the maximum cell density of the clone was 6.1 ${\times}$ 10$^{6}$ cells/ml at day three of the incubation. Based on the pattern of genomic DNA laddering and the microscopic observation of apoptotic bodies, the more apoptotic bodies are induced in Tn 5B1-4 control cell clone at higher agitation speed. This result shows that the shear stress can be a main factor in inducing apoptosis in spinner flask culture. At low agitation speed, cell clone expressing Bcl-2 was more effective in delaying the onset of apoptosis than the cell clone expressing P35. On the other hand, at high agitation speed, cell clones expressing baculovirus P35 was more effective in delaying the onset of apoptosis than the cell clone expressing Bcl-2. Therefore, anti-apoptotic genes, bcl-2 and baculovirus p35, can playa distinct role depending on agitation speed in the suspension culture.