• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.97-97
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• 2003

For many animals the centrosome consists of a pair of centrioles and surrounding pericentriolar materials (PCMs). PCMs have been known to play roles during cell division. It is known that centrioles are necessary to assemble centrosomal components. However, many types of oocytes undergo meiosis without centrioles. It is known that in nonmurine mammalian species, the sperm introduces an intact proximal centriole unlike sea urchin where two centrioles are introduced. In case of mouse sperm, the presence of centrosome is not clear In this study, a monoclonal antibody was developed to investigate centrosome during mouse germ cell and early embryo development. Results of immunostaining and Western blotting in CHO cells suggest that the monoclonal antibody recognizes a nuclear and centrosomal protein, thus called NCP. The NCP monoclonal antibody was used to screen a cDNA expression library prepared from 12.5 mouse brain to isolate NCP gene. Nucleotide size of NCP gene obtained from immunoscreening was about 5.5kb. It is determined that the NCP may be closely related with pericentriolar material -1 gene (Pcm-1) from the result of sequencing analysis. The molecular weight, 66kDa, calculated by known DNA sequence in database is consistent with that of detected from Western blotting using CHO cell lysates. Therefore, it is assumed that NCP may be alternative splicing form of Pcm-1 of which molecular weight is 228kDa. In mouse oocytes, NCP was distributed in nucleus as in CHO cells. It was shown that the NCP was localized around neck region, probably the centrosome in mouse neck region. Interestingly, dramatic change in distribution of NCP was also shown in male germ cell development. Finally, we observed the cellular distribution of NCP during early embryo development. NCP was detected in nucleus as well as centrosome foci. It is suggested that the centrioles reassembly we occurring in blastocysts and then affects the distribution of NCP.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.49-49
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• 2003

본 연구에서는 vesicular stomatitis virus G glycoprotein (VSV-G)를 envelope로 가지는 pantropic retrovirus vector system을 이용하여 재조합 human FSH 유전자가 전이된 형질전환 닭을 생산하고자 하였다. Human FSH $\alpha$ 및 $\beta$ 유전자와 CTP linker는 human pituitary gland cDNA library에서 RT-PCR 방법을 이용하여 cloning하였으며, 각각의 fragment는 FSH$\beta$-CTP-FSH$\alpha$ 순서의 단일사슬로 연결하였다. 연결된 FSH$\beta$-CTP-FSH$\alpha$는 retroviral vector 내의 $\beta$-actin promoter의 조절 하에 도입한 후, PT67 packaging cell line에 transfection하여 virus를 생산하였으며 생산된 virus는 pantropic한 virus producing cell인 GP293에 infection하여 FSH 유전자가 도입된 virus를 생산하였다. FSH 유전자의 발현을 in vitro에서 확인하기 위하여 CHO (chinese hamster ovary) 세포에 virus를 감염시킨 후, 세포의 배양액을 취하여 electrochemilumine-scence immunoassay 방법으로 정량하였다. In vitro에서 전이 후 발현이 확인된 FSH 외래유전자의 retroviral vector virus를 초원심분리로 고농축하여 stageX의 계란의 배반엽 층에 주입하였으며, 그 결과 18%의 부화율과 91%의 부화한 닭의 유전자 전이율을 확인할 수 있었다. 전이된 유전자의 확인은 FSH$\beta$와 Neo 유전자에 대한 primer를 이용한 RT-PCR의 방법을 이용하였다. In vitro에서와는 달리 in vivo에서는 FSH 유전자의 전이는 확인되었으나 발현을 확인하지는 못하였는데, 이는 적은 수의 실험군이 형질전환율에 비해 상대적이지 못하였거나, 외래 유전자인 FSH의 발현에 의한 생리적인 부작용이 유발되어 해당개체가 부화되지 못한 것으로 추정된다. 본 문제점을 해결하기 위하여 실험군의 수를 늘리고 외래 유전자에 대한 controllable expression system이 보완될 필요성이 요구되며, 이러한 점이 해결된다면 높은 유전자 전이율에 기인하여 retrovirus를 이용한 형질전환 방법은 형질전환 가금의 생산에 있어서 매우 효율적이고 주목할 만한 방법으로 사료된다.

• Korean Institute of Interior Design Journal
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• v.14 no.6 s.53
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• pp.177-184
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• 2005

The 21st Century Museum of Contemporary Art, Kanazawa as a regional public culture facility is the place where it is possible to think over the roles and meanings of a museum. The urn of this research is to provide information for exploring a desirable direction of the design of a regional museum by analyzing the architectural meanings and public property of the 21st Century Museum of Contemporary Art, Kanazawa. In addition, we arranged the totes and meanings of a regional museum and the concept of the public property in architecture. We also analyzed the architectural quality and the expression of the public property in this museum, as well as the value and meaning of a museum through field researches. The results of this research are as follows. First, this museum breaks with the existing stereotyped Idea to be a space open to the citizens. It secures a wide public space and provides the convenience of access and every facility for citizens. Second, the floor plan is divided into the interchanging area for citizens and an exhibition area; the outskirts of the inside of the museum are planned to be used free of charge for an interchanging space of citizens as a public-owned space of citizens. Third, the public-owned space of citizens includes a citizen exhibition hall, an art library, a kids studio and rest space, which ate all designed lot everyone to enjoy freely. Last, the exterior shape of the museum is simple but harmonize with the surroundings. It has an unique shape showing the local identity, and its most special properties are the convenience of access, introduction of a free space inside, and security of a huge space for children and citizens.

• Microbiology and Biotechnology Letters
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• v.27 no.6
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• pp.491-499
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• 1999

A great variety of the volatile metabolic by-products was formed in yeast cell during alcohol fermentation. The seibel grape (Vitis labrasca) which was grown in the Southern Korea used for wines. The objective of this research was to identify the volatile flavor compounds during alcohol fermentation and aging at 12$^{\circ}C$. saccharomyces cerevisiae and Schizosaccharomyces pombe were inoculated and fermented in seibel grape must. The volatile flavor compounds of logarithmic, stationary and death phases were extracted, concentrated and identified by gas chromatography/mass spectrometer (GC/MS). The volatile flavor compounds were determined by a Hewlett-Packard 5890 II Plus GC which was equipped with Supelcowax 10 fused silica capillary column (60m$\times$0.32mm$\times$0.25${\mu}{\textrm}{m}$ film thickness) wall coated with polyethyleneglycerol. The scan detection method allowed the comparison of the spectrum from the chromatogram of volatile flavor compounds to those in data Wileynbs base library. Among the volatile compounds collected by ether-hexane extraction method, the evolution of 20 main compounds, such as 9 esters (ethyl butyrate, isoamyl acetate, ethyl caproate, n-hexyl acetate, ethl caprylate, ethyl caprate, diethy succinate, ethyl hexadecanoate, 2-pheneethyl acetate), 4 alcohols (3-methyl-1-butanol, 1-hexanol, 1-heptanol, benzoethanol), 4 ketones and acids (2-octanone, caproic acid, caprylic acid, capric acid), 2 furan and phenol (2,6-bis(1,1-dimethyl ethyl)phenol, 2,3-dihydrobenzofuran) were observed during alcohol fermentation and aging. The production of the esters during alcohol fermentation with S. cerevisiae was higher than those of Sch. pombe. The sensory scores of the aged wine samples in aroma, taste and overall acceptability were not significantly different(p<0.05).

• Journal of Microbiology and Biotechnology
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• v.17 no.6
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• pp.891-896
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• 2007

We screened the Arabidopsis cDNA library to identify functional suppressors of AtBI-1, a gene that suppresses cell death induced by Bax gene expression in yeast. Cdf 3 encodes a 118-amino-acid protein with a molecular mass of 25 kDa. This protein has two uncharacterized domains at amino acids residues 5-64 and 74-117. In the present study, CDF3 was found to induce growth defects in yeast and arrested yeast growth, although the cell-growth defect was somewhat less than that of Bax. Its localization in the inner mitochondria was essential for suppression of yeast-cell proliferation. The morphological abnormality of the intracellular network, which is a hallmark of AtBI-1, was attenuated by Cdf3 expression.

• Journal of Microbiology and Biotechnology
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• v.23 no.8
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• pp.1047-1054
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• 2013

Witches' broom of lime is a disease caused by Candidatus Phytoplasma aurantifolia, which represents the most significant global threat to the production of lime trees (Citrus aurantifolia). Conventional disease management strategies have shown little success, and new approaches based on genetic engineering need to be considered. The expression of recombinant antibodies and fragments thereof in plant cells is a powerful approach that can be used to suppress plant pathogens. We have developed a single-chain variable fragment antibody (scFvIMP6) against the immunodominant membrane protein (IMP) of witches' broom phytoplasma and expressed it in different plant cell compartments. We isolated scFvIMP6 from a naïve scFv phage display library and expressed it in bacteria to demonstrate its binding activity against both recombinant IMP and intact phytoplasma cells. The expression of scFvIMP6 in plants was evaluated by transferring the scFvIMP6 cDNA to plant expression vectors featuring constitutive or phloem specific promoters in cassettes with or without secretion signals, therefore causing the protein to accumulate either in the cytosol or apoplast. All constructs were transiently expressed in Nicotiana benthamiana by agroinfiltration, and antibodies of the anticipated size were detected by immunoblotting. Plant-derived scFvIMP6 was purified by affinity chromatography, and specific binding to recombinant IMP was demonstrated by enzyme-linked immunosorbent assay. Our results indicate that scFvIMP6 binds with high activity and can be used for the detection of Ca. Phytoplasma aurantifolia and is also a suitable candidate for stable expression in lime trees to suppress witches' broom of lime.

• Journal of Microbiology and Biotechnology
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• v.19 no.2
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• pp.147-154
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• 2009

The carboxylesterase-encoding gene(bioHs) of a newly isolated strain, Serratia sp. SES-01, was cloned from the genomic DNA library by detecting formation of transparent halo around the colony on LB-tributyrin agar plates. The amino acid sequence of BioHs was highly similar to the members of the BioH enzyme family involved in the biotin biosynthetic pathway; it showed the highest similarity(91%) with that of Serratia proteamaculans. To compare BioHs with other BioH enzymes, the relatively well-known bioHe gene of E. coli was cloned with PCR. After we achieved high-level expression of soluble BioHs and BioHe through the exploration of different culture conditions, the purified BioHs and BioHe enzymes were characterized in terms of specificity, activity, and stability. BioHe was generally more robust to a change in temperature and pH and an addition of organic solvents than BioHs. The two enzymes exhibited a strong preference for carboxylesterase rather than for thioesterase and were optimal at relatively low temperatures($20-40^{\circ}C$) and alkaline pHs(7.5-9.0). The results in this study strongly suggested that both the BioHs and BioHe enzymes would be potential candidates for use as a carboxylesterase in many industrial applications.

• Journal of Microbiology and Biotechnology
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• v.27 no.8
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• pp.1379-1385
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• 2017

The content of taxol in the bark of yews is very low, and this is not affordable from the environmental point of view. Thus, it is a necessity to look for alternative sources of taxol production to solve its supply. Currently, a large portion of the taxol in the market comes from chemical semi-synthesis, but the semi-synthetic precursors such as baccatin III and 10-deacetyl-baccatin III are extracted from needles and twigs of yew trees. Taxol-producing fungi as a renewable resource is a very promising way to increase the scale of taxol production. Our group has obtained a taxol-producing endophytic fungus, Aspergillus niger subsp. taxi HD86-9, to examine if A. niger can produce the taxanes. Six compounds from the fermentation broth of strain HD86-9 were isolated and identified by $^1H$ NMR, $^{13}C$ NMR, and ESI-MS. The results showed that the six compounds included four taxane diterpenoids (taxol, cephalomannine, baccatin III, and 10-deacetyl-baccatin III) and two non-taxane compounds (${\beta}-sitosterol$ and flavonoid isovitexin). The study verified that the taxanes can be produced by the A. niger, which is very important to taxol production via chemical semi-synthesis. Additionally, the finding is potentially very significant to solve the taxol semi-synthetic precursors extracted from needles and twigs of yew trees, and the precursor production can be easily increased through the culture condition optimization, genetic breeding, and metabolic engineering of the A. niger.

• Journal of the Institute of Electronics Engineers of Korea SD
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• v.47 no.12
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• pp.48-54
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• 2010

This paper presents a high-performance architecture of integer-pel motion estimation circuit for H.264 video CODEC. Full search algorithm guarantees the best results by examining all candidate blocks. However, the full search algorithm requires a huge amount of computation and data. Many fast search algorithms have been proposed to reduce the computational efforts. The disadvantage of these algorithms is that data access from or to memory is very irregular and data reuse is difficult. In this paper, we propose an efficient integer-pixel motion estimation algorithm and the circuit architecture to improve the processing speed and reduce the external memory bandwidth. The proposed circuit supports seven kinds of variable block sizes and generates 41 motion vectors. We described the proposed high-performance motion estimation circuit at R1L and verified its operation on FPGA board. The circuit synthesized by using l30nm CMOS standard cell library processes 139.8 1080HD ($1,920{\times}1,088$) image frames per second and supports up to H.264 level 5.1.

• KSBB Journal
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• v.16 no.6
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• pp.562-567
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• 2001

Aspergillus niger LK harboring the enantioselective epoxide hydrolase (EHase) activity was isolated, and enantioselectivity of EHase was tested for various racemic aromatic epoxides. The gene encoding epoxide hydrolase was cloned from cDNA library generated by reverse transcriptase-polymerase chain reaction of the isolated total mRNA. Sequence analysis showed that the cloned gene encodes 398 amino acids with a deduced molecular mass of 44.5 kDa. Database comparison of the amino acid sequence reveals that it is similar to fungal EHase, whereas the sequence identity with bacterial EHase is very low. Recombinant expression of the cloned EHase in Escherichia coli BL21 yielded an active EHases, which can offer a potential biocatalyst for the production of chiral epoxides.