• Journal of Microbiology and Biotechnology
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• v.32 no.5
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• pp.621-629
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• 2022

Bacterial outer membrane vesicles (OMVs) typically contain multiple immunogenic molecules that include antigenic proteins, making them good candidates for vaccine development. In animal models, vaccination with OMVs has been shown to confer protective immune responses against many bacterial diseases. It is possible to genetically introduce heterologous protein antigens to the bacterial host that can then be produced and relocated to reside within the OMVs by means of the host secretion mechanisms. Accordingly, in this study we sought to develop a novel platform for recombinant OMV (rOMV) production in the widely used bacterial expression host species, Escherichia coli. Three different lipoprotein signal peptides including their Lol signals and tether sequences-from Neisseria meningitidis fHbp, Leptospira interrogans LipL32, and Campylobactor jejuni JlpA-were combined upstream to the GFPmut2 model protein, resulting in three recombinant plasmids. Pilot expression studies showed that the fusion between fHbp and GFPmut2 was the only promising construct; therefore, we used this construct for large-scale expression. After inducing recombinant protein expression, the nanovesicles were harvested from cell-free culture media by ultrafiltration and ultracentrifugation. Transmission electron microscopy demonstrated that the obtained rOMVs were closed, circular single-membrane particles, 20-200 nm in size. Western blotting confirmed the presence of GFPmut2 in the isolated vesicles. Collectively, although this is a non-optimized, proof-of-concept study, it demonstrates the feasibility of this platform in directing target proteins into the vesicles for OMV-based vaccine development.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.213-221
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• 2001

This study was conducted to examine in vitro developmental ability of porcine embryos after somatic cell nuclear transfer. The porcine ear fell was cultured in vitro for confluency in serum-starvation condition(TCM-199 + 0.5% FBS) far 3~6 days of cell confluency. The zona pellucida of IVM oocytes were partially drilled using laser system. Single somatic cell was individually transferred into enucleated oocytes. And the reconstructed embryos were electrically fused(single DC 1.9kv/cm, 30$\mu$ sec) with 0.3M mannitol. After electrofusion, embryos were activated(single AC 5v/mm, 5sec) and cultured in HCSU-23 medium containing 10% FBS at 39$^{\circ}C$, 5% $CO_2$ in air for 6 to 8 days. The fusion rate of donor cells was 45.6, 36.8 and 46.1% in 3~4, 5~6 days of serum starvation and non serum starvation(N-S), and were 52.7. 53.0 and 51.7% in 1~2. 5~6 and 13~14 passages of donor cell culture, respectively. No significant difference was found in the fusion rate of donor cells by the duration of serum starvation treatment or the number of donor cell passages. By the size of donor cells, however, the fusion rate was significantly higher(P<0.05) for reconstructed embryos derived from 25r $\mu$m $\geq$ site of donor cells (65.3%) than that of 25~30$\mu$ m(42.5%) or 30$\mu$ m(45.5%)$\leq$ cells. The cleavage rate was significantly (P<0.05) higher in 3~4 darts of serum starvation treatment(67.1%) than that in N-S (50.7%) or 5~6 days of starvation(57.1%). The activation rate by the size of donor cells in fused oocytes was 56.5, 68.8 and 58.5%, respectively, and was not significant.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.89-95
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• 2007

The present study was conducted to examine some factors affecting in vitro development and fecundity of embryos recloned with somatic cell nuclear transfer (SCNT). Fibroblast cells retrieved from the ear of a 3-week-old, cloned Korean goat (Jinsoonny) were used as karyoplast donors and serum-starvation was conducted in tissue culture medium (TCM)-199 supplemented with 0.5% FBS. Recipient oocytes were surgically collected by flushing the oviducts 35 h after hCG injection following FSH priming. The zonae pellucidae of the oocytes were partially perforated with a laser drill and a donor cell was transferred into an enucleated oocyte. The couplets were electrically fused and activated by ionomycin (5 min) and 6-DMAP (4 h). The reconstructed embryos were cultured in mSOF medium containing 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $%O_2$, 90% $N_2$ for 12 to 15 h. Re-cloned embryos (2- to 4-cell stages) were surgically transferred into the oviducts of the recipients and pregnancy was subsequently diagnosed by progesterone assay and ultrasound on Days 21 and 63 of pregnancy. The fusion rate following 1st fusion pulse was higher (p<0.05) in 2nd cloning (65.9%) compared to 1st cloning (51.0%), but it was not different in the other groups. The rate of cleavage after fusion was significantly higher (p<0.05) in 1st (77.7%) than in 2nd cloning (56.0%). A total of 175 re-cloned embryos were transferred into 28 recipients. On day 21 and 60 after transfer, 11 (39.3%) and 4 recipients (17.4%) were pregnancy, respectively. In comparison of pregnancy rate by estrous synchronization, a total of 66 and 109 re-cloned embryos were transferred into 11 recipients in natural estrus and 17 recipients in induced estrus, respectively. Five (45.4%) and 2 recipients (18.2%) in natural estrus were pregnant on days 21 and 63 while 6 (35.3%) and 2 (11.8%) recipients in induced estrus were pregnant, respectively. These results show that recloning of goat can be achieved by SCNT and estrous synchronization between donor and recipient animals may be one of the major factors affecting success rate.

• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.293-302
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• 1997

Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

• KSBB Journal
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• v.19 no.1
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• pp.17-26
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• 2004

In this study the extracellular production of 5-aminolevulinic aicd (ALA) by recombinant E. coli BL2l (DE3) pLysS harboring the plasmid pFLS45 are investigated. Optimum concentrations of succinic acid and glycine for cell growth and ALA production were found to be 30 mM and 15 mM, respectively. Levulinic acid (LA) as an inhibitor of ALAD was added to the culture medium in the end of exponential cell growth phase and its optimum concentration was 30 mM. Growth of recombinant E. coli BL2l (DE3) pLysS (pFLS45) was largely dependent upon the pH value of culture medium. When the pH of culture medium was in the range of 6.0 and 6.5, high cell mass and ALA production were obtained. IPTG induction for the expression of the fusion gene did not enhance the production of ALA. Recombinant cell grew at 30't faster than at 37$^{\circ}C$, but ALA productivity was lower than at 37$^{\circ}C$. Repeated addition of glycine, succinic acid, and LA increased the production of ALA and the inhibition of intracellular ALA dehydratase activity, with up to 1.3 g/L ALA having been produced in the cultivation.

• Science of Emotion and Sensibility
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• v.15 no.1
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• pp.121-132
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• 2012

Recently, interest in emotional technology has escalated and its application has extended to many fields, from both an industrial (product) and service (culture, tourism, medical, education, advertising) perspective. Moreover, culture has become a fundamental aspect of modern industry, playing key roles in: gaming, exhibitions, performances, sports, tourism, design, edutainment, as well as various content distribution industries. The prospect of applying emotional technology for cultural content industries makes up for more than half of the 'plan-manufacture-distribution-marketing' process, and thereby also serves as driving force for the growth of a nation. The primary objective of the following research is analyze successful cases from the past through utilization of emotional technology, and to speculate on efficient directions for future research into developing emotional contents. To achieve this, some of the key terms have been defined and elaborated for the sake of clarity. The terms are as follows: emotion, emotion engineering, science of emotion, emotional technology, and emotional contents. Furthermore, studies were conducted based on the six different fields surrounding CT R&D to observe how projects involving emotional technology have succeeded both on a national and global scale. Based on this analysis, this research aims to develop personalized 'Concierge' service-providing contents, contents designed to maximize performance ability of humans, and contents that could be controlled simply via emotion to effectively spread the culture of Korea by focussing on 'fusion' content development. The following research data may hopefully serve as a basic reference to industries navigating towards emotional content development.

• Korean Journal of Heritage: History & Science
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• v.54 no.3
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• pp.204-227
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• 2021

This paper proposed the validity and necessity of establishing the Jungwon Historical and Cultural Area by reviewing the historical characteristics of the name Jungwon and its temporal and spatial range. We also explored ways to utilize the Jungwon historical and cultural Area under the Special Act on the Maintenance of Historical and Cultural Areas. Jungwon, which means the center of the country, has strategic value as a key area connecting the Korean Peninsula to the north and south through the Geripryeong and Jukryeong. It also includes Chungju, which connects the Namhan and Bukhan Rivers . From the Mahan period, the areas of Cheongju and Jincheon, the Miho Stream basin to the west, and Chungju to the east were closely connected to each other. Afterwards, Baekje, Koguryo, and Silla engaged in a fierce battle to occupy the Jungwon area. Jungwon culture was born through the negotiation and fusion of the cultures of the three kingdoms. In particular, Silla continued to value the Jungwon area, demonstrated by the installation of Seowongyeong in Cheongju and Bukwongyeong in Wonju, centering on the Jungwongyeong. Since we have a historical view with a focus on ancient kingdoms, we have overlooked the full value of the history and culture of Jungwon. We need to recognize the importance of Jungwon area and think about how to utilize it. Therefore, we suggested in this paper that there be more public information services on the cultural heritage of Jungwon, the historical heritage experience, and the education programs for local residents. The development and operation of Jungwon's history, culture, and tourism contents, as well as the various academic research papers should be used to reveal the identity of the Jungwon historical and cultural area.

• The Journal of the Convergence on Culture Technology
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• v.9 no.3
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• pp.399-406
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• 2023

Due to the increasing influence of the Korean Wave, traditional themes are emerging to the forefront of performance content development, emphasizing the necessity of applying the art of storytelling to the creative process. The purpose of this research is to examine the differentiated costume design of the Pansori ballet "Dokkaeba! Dokkaeba!," developed through the implementation of storytelling in its performance creation process. Choreographers, writers, and costume designer collaborated throughout the storytelling process from planning to content creation. This resulted in a clarified depiction of the Dokkaebi as divine beings with transcendental powers who are simultaneously friendly and familiar entities, often appearing in the form of humans and objects. Accordingly, the costume design based on this collaborative storytelling deliberately avoided the appearance of Dokkaebi reminiscent of 'Kwimyeonwa' or Japanese 'Oni', typically expressed with horns or decorations, and instead implemented novel design elements, such as fusion Hanbok with a variety of dimensionally aesthetic accessories, meaningful color contrast in modern clothing, textural elements indirectly expressing character and transmogrification, and daily clothing attire to illustrate the humanized Dokkaebi. Therefore, this study seeks to contribute to the production of new performance art showcasing the Dokkaebi by identifying the diversity of expression and direction within costume design.

• (The) Research of the performance art and culture
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• no.32
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• pp.399-426
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• 2016

Sanjo has a quintessential value not only because of its musical depth but also because of its style of music that best expresses Korean sentiment. However, new values are being established from different viewpoints as time passes so Sanjo's values need a modification in order to encompass a contemporary value that is required and accepted by modern society. In this context, while focusing on communication with the public, I contemplated the developmental direction of Sanjo in five perspectives; The Social realization of value, The Experimental and social transformation, The Social diffusion of creation, The Leap of fusion and harmony and finally The Socialization of contents. In the perspective of 'social realization of value', Sanjo refers to creative activity as a new 'duneum' which allows traditional and creative activity to deviate freely while still being under the Sanjo guideline. Either way, it has a periodical value because new forms with new rhythms are the only ways to communicate with the modern public. When these values can be understood by modern society can Sanjo be revived and be acknowledged as an infinite value. Secondly as an experiment and social transformation, there is a transformation of musical instruments in the 21th century. Our musical instruments should be transformed to effectively perform our music rather than to perform Western music. Third, social diffusion of creation should be the 'new Sanjo festival in 21th century' which can facilitate the communication with the public. Fourth, regarding leap of fusion and harmony, I suggest a performing culture consisting of 'Storytelling Sanjo' and 'Media Art' which will ceaselessly evolve Sanjo performance as a medium to communicate with the public. Finally, in regards to the socialization of contents, I emphasize that Sanjo should have contents of mass media as a way of means to help utilize mass media.

• Korean Journal of Animal Reproduction
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• v.22 no.4
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• pp.425-435
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• 1998

To improve the efficiency of nuclear transplantation in bovine, in this study the development in vitro of nuclear transferred (NT) embryos was compared by different activation regimens of the enucleated oocytes. The effect of developmental stage and culture system of donor nuclei on fusion and development in vitro of NT embryos were also evaluated. Oocytes were collected from Hanwoo ovaries obtained from slaughterhouse and matured in Ham's F-10 supplemented with hormones. After 20~22 h maturation, the oocytes were vortexed to be free from cumulus cells and subsequently their nucleus and the first polar body were removed. Enucleated oocytes were divided into 3 groups for activation; the oocytes of group I were activated with ionomycin for 5 min and subsequently incubated in 6-dimetylarninopurine (DMAP) for 4 h, Those of group II were treated with DMAP for 4 h at 39 h after onset of in vitro maturation (IVM) and those of group III were kept in room temperature ($25^{\circ}C$) for 3 h at 39 h after onset of IVM. After in vitro fertilization (IVF) the embryos for muclear donor were cultured either by group culture (20 embryos /50 ${mu}ell$ drop) or individually (1 embryo /50 ${mu}ell$ drop) for 4 day and 5 day. At day 4 and 5 after IVF, blastomeres were separated in calcium-magnesium free medium, and then classified into small (day 5: $\leq$ 38 ${\mu}{\textrm}{m}$, day 4: $\leq$ 46 ${\mu}{\textrm}{m}$) and large (day 5 : $\geq$ 38 ${\mu}{\textrm}{m}$, day 4 ; $\geq$ 46 ${\mu}{\textrm}{m}$). The separated blastomeres were replaced into enucleated and activated recipient cytoplasm. The blastomere-oocyte complexes were fused by electrically. The NT embryos were cultured in TCM-199 containing 10% FCS in 39$^{\circ}C$, 5% $CO_2$ incubator for 7 day. The results obtained were summarized as follows; There were no differences in fusion and development to blastocyst between groups as group I (68%, 10%), group II (75%, 14%) and group III (73%, 9%), respectively. However, the cell number in blastocyst of NT embryos in group III were significantly fewer than in the other groups (P＜0.05). No differences in fusion and development to blastocyst were found between individual or group cultured and between small or large blastomeres of day 4 and day 5 donor embryos. From these results, it was concluded that the combination of ionomycin and DMAP, or treatment of DMAP at 39 h after onset of IVM were useful for the efficient of production of NT bovine embryos, and the individual cultured embryos could be simply used as donor nuclei for NT bovine embryo.