• Molecules and Cells
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• v.41 no.4
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• pp.342-350
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• 2018

Flowering time is determined by florigens. These genes include, Heading date 3a (Hd3a) and Rice FT 1 (RFT1) in rice, which are specifically expressed in the vascular tissues of leaves at the floral transition stage. To study the cis-regulatory elements present in the promoter region of Hd3a, we generated transgenic plants carrying the 1.75-kb promoter fragment of Hd3a that was fused to the ${\beta}$-glucuronidase (GUS) reporter gene. Plants expressing this construct conferred a vascular cell-specific expression pattern for the reporter gene. However, GUS was expressed in leaves at all developmental stages, including the early seedling stage when Hd3a was not detected. Furthermore, the reporter was expressed in roots at all stages. This suggests that the 1.75-kb region lackings cis-elements that regulate leaf-specific expression at the appropriate developmental stages. Deletion analyses of the promoter region indicated that regulatory elements determining vascular cell-specific expression are present in the 200-bp region between -245 bp and -45 bp from the transcription initiation site. By transforming the Hd3a-GUS construct to rice cultivar 'Taichung 65' which is defective in Ehd1, we observed that Ehd1 is the major regulatory element that controls Hd3a promoter activity.

• Microbiology and Biotechnology Letters
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• v.30 no.3
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• pp.206-209
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• 2002

For the expression of CGTase gene(cgtS) kom Bacillus stearothemophilus NO2 in Saccharomyces cerevisiae, cgtS gene was subcloned into the Eschepichia coll-yeast shuttle vector, pVT103-U. The constructed plasmid, pVT-CGTS was introduced to 5. cemi-siae 2805 cell, and then the cgtS gene under the control of adhl promoter was successfully expressed in the yeast transformant and 87% of the total activity was detected into the fermentation medium. Therefore, the signal peptide of B. stearothemephilus NO2 CeTase showed high secretion efficiency in 5. cerevisiae. Optimal conditions of the recombinant yeast cell f3r expression of CGTase was achieved, when 5. cerevisiae 2805/pv7-CGTS was cultivated on YP medium at 2% dextrose, pH 5.5,$30^{\circ}C$ and the expression level of CGTase was 0.624units/mL for 48 h culture.

• The Korean Journal of Food And Nutrition
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• v.25 no.1
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• pp.181-187
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• 2012

Grifola frondosa has been used as an herbal medicine for the treatment of cancer, diabetes mellitus and high blood pressure. In this study, functional polysaccharide was obtained from Grifola frondosa using four different extraction methods: hot water(HwFP), homogenize(HgFP), acid(AcFP), and alkali(AlFP) extraction methods. The effects of these extracts on KB and HepG2 cell lines were then examined for any anti-cancer activity. Alkaline extraction produced a yield of 0.175% and the total sugar content of the extract was 54.97%. We were able to confirm that the polysaccharide extracts from the mushroom produce an anti-cancer effect. The cytotoxicity of AlFP and AcFP against HepG2 cells were 22.86% and 28.88%, respectively, and the cytotoxicity of AlFP against the KB cell lines was 47.76% at a concentration of 1,000 ${\mu}g/m{\ell}$. Therefore, these results suggest that the optimum method for extracting functional polysaccharides from G. frondosa is the alkali extraction method.

• Korean Journal of Animal Reproduction
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• v.18 no.3
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• pp.199-206
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• 1994

We present here a new PCR-based cloning technique that allows the different PCR products during mouse embryogenesis. Recently, mRNA differential display described by Liang & Pardee (Science 257, 1992) and re-confirmed by Zimermann & Schultz (PNAS 91,1994). This method will detect the appropriate changes in the temporal patterns of expression or in the transition from maternal control to zygotic control as well as the functional difference of embryo with polyspermy or monospermy, the difference of expression between successfully hatched blastocyst and blastocyst failed to hatching, response to agents, and cell cycle regulation. By this methods, we have cloned an eDNA, which showed mouse 2 cell specific expression. Genomic DNA digested with EcoRI showed approximately 15 kb and then showed higher expression in fetal liver rather than adult liver. Furthermore, this gene is likely to have 2 mRNA by alternative splicing.

• Journal of the Korea Institute of Information and Communication Engineering
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• v.20 no.2
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• pp.317-326
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• 2016

In this paper, a logic eFuse (electrical Fuse) OTP (One-Time Programmable) memory IP (Intellectual Property) using only logic transistors to reduce the development cost and period of OTP memory IPs is designed. To secure the reliability of other IPs than the OTP memory IP, a higher voltage of 2,4V than VDD (=1.5V) is supplied to only eFuse links of eFuse OTP memory cells directly through an external pad FSOURCE coming from test equipment in testing wafers. Also, an eFuse OTP memory cell of which power is supplied through FSOURCE and hence the program power is increased in a two-dimensional memory array of 128 rows by 8 columns being also able to make the decoding logic implemented in small area. The layout size of the designed 1kb eFuse OTP memory IP with the Dongbu HiTek's 110nm CIS process is $295.595{\mu}m{\times}455.873{\mu}m$ ($=0.134mm^2$).

• Journal of Dairy Science and Biotechnology
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• v.21 no.2
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• pp.114-119
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• 2003

Acidocin 4A produced by Lactobacillus acidophilus GP4A was purified to homogeneity by ammonium sulfate precipitation and sequential chromatographies containing Octyl sepharose CL-4B column, $C_{18}$ Sep-Pak Cartridge, $C_{18}$ RP HPLC and HPLC gel filtration. Tricine SDS-PACE resulted in a single band with estimated molecular mass of 4.1 kDa corresponding to the polypeptide weight marker. Electron microscopy of acidocin-treated indicator cells(L. delbrueckii subsp. lactis ATCC 4797) confirmed that acidocin 4A presented bacteriolytic effect, resulting in cell lysis. Curing trial using ethidium bromide (EtBr) was carried out to examine whether acidocin 4A determinant was encoded either by chromosome or on plasmid. The plasmid designated as pLA4A, being about 20 kb in size, was responsible for acidocin 4A production and immunity to host cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.4
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• pp.669-674
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• 1997

This study was attempted to express human lactoferrin gene that has importance as a functional additive in food industry. Lactoferrin has distinctive antibacterial properties. Also, a number of phy-siological roles have been postulated for the lactoferrin in the modulation of immune and inflamatory responses and as a growth factor. Since it did not show feasible growth inhibition by antimicrobial test against HLF, Pichia pastoris was selected the best lactoferrin expression host. HLF expression plasmid pHIL-SI was integrated into the genomic DNA of P. pastoris GSl15. The integration was confirmed not only with 2.4Kb fragment of HLF gene by PCR(polymerase chain reaction) product, but also with same size of specific signal by southern blotting. Among the various pichica transformants, the JY-1 cell showed a positive response for the expression of HLF by the immunoblotting anaysis. The recombinant HLF protein was started to be secreated at 48hr of culture and reached at the highest secreation level at 96hr.

• Restorative Dentistry and Endodontics
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• v.21 no.1
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• pp.403-424
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• 1996

Cell culture methods have been used to assess the cytotoxicity of dental materials. Different paramaters are used to monitor cytotoxic effects. But it is difficult to compare each investigator's results with different methods. The objective of this study was to investigate cytotoxic effect of several retrograde filling materials according to cell lines and assay methods. Cytotoxicity of Bestalloy (Dogmyung, Korea), Prisma APH(Densply International Inc., U.S.A.), Clearfil FII (Kuraray Co., Japan), Fuji II (GC Co., Japan), Fuji II LC (GC Co., Japan) and IRM (Densply Co., U.S.A.) on L929, 3T3 and KB permanent cell lines was measured. Radiochromium, Lactate dehydrogenase (LDH) release method and colorimetric assays, namely neutral red (NR) and MTT were used. Each material was mixed according to the manufacturer's instruction. They were tested as solid and extracted state. Cell culture media were added to each mixed or solid materials then the solution was collected and used as extract solutions. Solid Fuji II showed mild cytotoxicity on three cell lines using radiochromium release method. There was no difference in cytotoxicity of extract solution group using radiochromium release method. In colorimetric assay immediate Fuji II group and all the IRM groups showed severe cytotoxic effect. Difference in cyctotoxicity was due to rather kinds of cell lines than assay methods. Solid Fuji II and IRM showed mild cytotoxicity on three cell lines. But extract solutions had different cytotoxic effect according to cell lines using LDH release assay. Light-cured glass ionomer had mild to moderate degree of cytotoxicity on three cell lines. Cytotoxicity was affected by specimen prepaton. Susceptibility of each cell ines were also affected by assay emthods. It was suggested that cytotoxicity study using only one cell line and/or assay method might not accurately reflect the real toxic nature of dental biomaterials.

• Food Science of Animal Resources
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• v.22 no.4
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• pp.348-351
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• 2002

This study was carried out to investigate the anticancer effect of extracts from sulfur fed duck carcass. Growth inhibition of cancer cell lines was measured by MTT assay. Eleven cancer cell lines, such as Calu-3(human lung carcinoma), SK-MES-1(human lung carcinoma), HL6O(human leukemia), KB(human epidermoid of mouth carcinoma), Farrow(human melanoma), HEP-2(human larynx carcinoma), SNU-1(human stomach carcinoma), K-562 (human leukemia), WiDr(human colon carcinoma), P388(mouse leukemia) and 3LL(mouse lung carcinoma) showed the growth inhibition higher than 50%, but those, such as SF-188(human brain carcinoma), A-549(human lung carcinoma) and HEC-lB(human uterus carcinoma) showed the growth inhibition lower than 50% in the extract of sulfur fed duck carcass at the concentration of 10 mg/㎖. The sulfur fed duck carcass extract had better growth inhibition than the normal counterpart against various cancer cell lines at the concentration of 10 mg/㎖. When the effect of growth inhibition of an effluent by different concentrations of methyl alcohol(25, 50, 75 and 100%) tested on Diaion HP-20 column chromatography, an effluent by concentration of 100% methyl alcohol showed the most strong effect of growth inhibition against HEP-2(human larynx carcinoma).

• International Journal of Oral Biology
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• v.34 no.1
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• pp.21-28
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• 2009

Mutations in DLX3 are associated with both autosomal dominant hypoplastic hypomaturation amelogenesis imperfecta (ADHHAI) and tricho-dento-osseous (TDO) syndrome. ADHHAI is caused by a c.561_562delCT (2bp-del DLX3) mutation whereas TDO syndrome is associated with a c.571_574delGGGG (4bp-del DLX3) mutation. However, although the causal relationships between DLX3 and an enamel phenotype have been established, the pathophysiological role of DLX3 mutations in enamel development has not yet been clarified. In our current study, we prepared expression vectors for wild type and deletion mutant DLX3 products (4bp-del DLX3, 2bp-del DLX3) and examined the effects of their overexpression on the expression of the enamel matrix proteins and proteases. Wild type DLX3 enhanced the expression of matrix metalloprotease 20 (MMP20) mRNA and protein in murine ameloblast-like cells. However, neither a 4bp-del nor 2bp-del DLX3 increased MMP20 expression. Wild type DLX3, but not the above DLX3 mutants, also increased the activity of reporters containing 1.5 kb or 0.5 kb of the MMP20 promoter. An examination of protein stability showed that the half-life of wild type DLX3 protein was less than 12 h whilst that of both deletion mutants was longer than 24 h. Endogenous Dlx3 was also found to be continuously expressed during ameloblast differentiation. Since inactivating mutations in the gene encoding MMP20 are associated with amelogenesis imperfecta, the inability of 4bp-del or 2bp-del DLX3 to induce MMP20 expression suggests a possible involvement of such mutations in the enamel phenotype associated with TDO syndrome or ADHHAI.