• Development and Reproduction
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• v.7 no.2
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• pp.127-136
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• 2003

The present study was conducted to investigate gene expression profile of mouse ovaries during the primordial-primary follicle transition. We isolated total RNA from mouse ovaries at day1(contains only primordial follicles) and day5(contains both primordial and primary follicles) and synthesized cDNA using annealing control primers(ACP, Seegene, Inc., Seoul, Korea). Using 80 different ACPs for PCR, we cloned, sequenced, and analyzed identities of 41 differentially expressed genes(DEGs). According to BLAST analysis, sequences of 33 clones significantly matched database entries, 4 clones were novel, and 4 clones were ESTs. We selected 8 DEGs with interesting functions, Anx11 and Pepp2-Pending highly expressed in day1 ovary, while Apg3/Autlp-like, BPOZ, Ches1, Kcmf1, NHE3, Nid2, Ninj1, SENP3, Suil-rsl, and TIAP/m-survivin highly expressed in days ovary, and confirmed their different expression between day1 ovaries and days ovaries using semi-quantitative RT-PCR. There was no false positive result. Using in situ hybridization, we found that almost all of genes studied were expressed in the oocyte from primordial follicle stage but expression decreased from primary follicle stage. Meanwhile their expression was increased in cuboidal granulosa cells. Different expression of BPOZ and TIAP/m-survivin between primordial and primary follicles was confirmed by using laser capture microdissection followed by real-time PCR BPOZ and TIAP/m-survivin expressed 4.5 and 3.4 fold higher in primary than primordial follicles, respectively. List of genes obtained from the present study will provide insights for the study of mechanism regulating primordial-primary follicle transition.

• Journal of the Korea Concrete Institute
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• v.27 no.6
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• pp.687-693
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• 2015

The present study proposed a k-value to determine the water-to-binder ratio of concrete using fly ash (FA) or ground granulated blast-furnace slag (GGBS) as a partial replacement of ordinary portland cement (OPC) with regard to an equivalent strength of OPC concrete. From the regression analysis using an extensive database including 7076 concrete mixes, k-values were determined for various water-to-binder ratios when the replacement ratio of OPC by the addition of FA or GGBS were below 50%. For deriving an equation to identify k-value, the relationship of concrete compressive strength and water-to-binder ratio was generalized by an exponential function. In general, k-values decreased with the increases in the addition of FA or GGBS for replacement of OPC and water-to-binder ratio. The rate in decreasing k-value against water-to-binder ratio was marginally affected by the addition of FA or GGBS, although a higher k-value was commonly obtained for GGBS concrete than for FA concrete at the same water-to-binder ratio. Consequently, the determined k-values were simplified as a function of water-to-binder ratio and the addition ratio of FA or GGBS as replacement of OPC.

• Journal of the Korea Concrete Institute
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• v.24 no.2
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• pp.137-145
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• 2012

Strength model for blasted furnace slag mortar blended with sodium was investigated in this study. The main parameters of AAS (alkali activated slag) mortar were dosage of alkali activator, water to binder ratio (W/B), and aggregate to binder ratio (A/B). For evaluating the property related to the dosage of alkali activator, sodium carbonate ($Na_2CO_3$) of 4~8% was added to 4% dosage of sodium hydroxide (NaOH). W/B and A/B was varied 0.45~0.60 and 2.05~2.85, respectively. An alkali quality coefficient combining the amounts of main compositions of source materials and sodium oxide ($Na_2O$) in sodium hydroxide and sodium carbonate is proposed to assess the compressive strength of alkali activated mortars. Test results clearly showed that the compressive strength development of alkali-activated mortars were significantly dependent on the proposed alkali quality coefficient. Compressive strength development of AAS mortars were also estimated using the formula specified in the previous study, which was calibrated using the collected database. Predictions from the simplified equations showed good agreements with the test results.

• Korean Journal of Breeding Science
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• v.43 no.5
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• pp.448-456
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• 2011

We used suppression subtractive hybridization (SSH) combined with mirror orientation selection (MOS) method to screen differentially expressed genes from red-fleshed kiwifruit 'Hongyang'. As a result, the 288 clones were obtained by subcloning PCR product and 192 clones that showed positive clones on colony PCR analysis were selected. All the positive clones were sequenced. After comparisons with the NCBI/Genbank database using the BLAST search revealed that 30 clones showed sequence similarity to genes from other organisms; 10 clones showed significant sequence similarity to known genes. Among these clones, 3 clones (AcF21, AcF42 and AcF106) had sequence homology to 1-aminicyclopropane-carboxylic acid (ACC)-oxidase (ACO) that known to be related to fruit ripening. The expression patterns of differentially expressed genes were further investigated to validate the SSH data by reverse transcription PCR (RT-PCR) and quantitative real-time PCR (qReal-time PCR) analysis. All the data from qReal-time PCR analysis coincide with the results obtained from RT-PCR analysis. Three clones were expressed at higher levels in 'Hongyang' than 'Hayward'. AcF21 was highly expressed in the other genes at 120 days after full bloom (DAFB) and 160 DAFB of 'Hongyang'.

• Journal of Life Science
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• v.29 no.6
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• pp.631-636
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• 2019

A new nitric oxide-induced (NOI) gene was isolated by screening ESTs from a cDNA library of dehydration-treated fibrous roots of sweetpotato (Ipomoea batatas). The 720 bp cDNA fragment, IbNOI, was sequenced, from which a 77 amino acid residue protein was deduced. A search of the protein BLAST database identified significant similarity to other plant NOI protein sequences. Quantitative RT-PCR analysis revealed diverse expression patterns of IbNOI in various tissues of the intact sweetpotato plant, and in leaves exposed to different stresses. The IbNOI gene was highly expressed in storage roots and suspension-cultured cells. In leaf tissues, IbNOI showed strong expression during sodium nitroprusside (SNP)-induced NO accumulation and chemical stress treatments. Expression of IbNOI was also induced under various abiotic stress conditions, such as dehydration, salt, and bacterial pathogen infection. These results suggest that IbNOI is involved in plant responses to diverse abiotic stresses and pathogen infection through a NO-related pathway.

• Horticultural Science & Technology
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• v.29 no.6
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• pp.633-640
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• 2011

Brassica rapa is an A genome model species for Brassica crop genetics, genomics, and breeding. With the completion of sequencing the B. rapa genome, functional analysis of the genome is forthcoming issue. The expressed sequence tags are fundamental resources supporting annotation and functional analysis of the genome including identification of tissue-specific genes and promoters. As of July 2011, 147,217 ESTs from 39 cDNA libraries of B. rapa are reported in the public database. However, little information can be retrieved from the sequences due to lack of organized databases. To leverage the sequence information and to maximize the use of publicly-available EST collections, the Brassica rapa tissue-specific EST database (BrTED) is developed. BrTED includes sequence information of 23,962 unigenes assembled by StackPack program. The unigene set is used as a query unit for various analyses such as BLAST against TAIR gene model, functional annotation using MIPS and UniProt, gene ontology analysis, and prediction of tissue-specific unigene sets based on statistics test. The database is composed of two main units, EST sequence processing and information retrieving unit and tissue-specific expression profile analysis unit. Information and data in both units are tightly inter-connected to each other using a web based browsing system. RT-PCR evaluation of 29 selected unigene sets successfully amplified amplicons from the target tissues of B. rapa. BrTED provided here allows the user to identify and analyze the expression of genes of interest and aid efforts to interpret the B. rapa genome through functional genomics. In addition, it can be used as a public resource in providing reference information to study the genus Brassica and other closely related crop crucifer plants.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.3
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• pp.449-456
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• 2018

Objective: Nanog, a homeodomain protein, has been investigated in humans and mice using embryonic stem cells (ESCs). Because of the limited availability of ESCs, few studies have reported the function and role of Nanog in porcine ESCs. Therefore, in this study, we investigated the location of the porcine Nanog chromosome and its basal promoter activity, which might have potential applications in development of ESCs specific marker as well as understanding its operating systems in the porcine. Methods: To characterize the porcine Nanog promoter, the 5'-flanking region of Nanog was isolated from cells of mini-pig ears. BLAST database search showed that there are two porcine Nanog genomic loci, chromosome 1 and 5, both of which contain an exon with a start codon. Deletion mutants from the 5'-flanking region of both loci were measured using the Dual-Luciferase Reporter Assay System, and a fluorescence marker, green fluorescence protein. Results: Promoter activity was detected in the sequences of chromosome 5, but not in those of chromosome 1. We identified the sequences from -99 to +194 that possessed promoter activity and contained transcription factor binding sites from deletion fragment analysis. Among the transcription factor binding sites, a Sp1 was found to play a crucial role in basal promoter activity, and point mutation of this site abolished its activity, confirming its role in promoter activity. Furthermore, gel shift analysis and chromatin immunoprecipitation analysis confirmed that Sp1 transcription factor binds to the Sp1 binding site in the porcine Nanog promoter. Taken together, these results show that Sp1 transcription factor is an essential element for porcine Nanog basal activity the same as in human and mouse. Conclusion: We showed that the porcine Nanog gene is located on porcine chromosome 5 and its basal transcriptional activity is controlled by Sp1 transcription factor.

• Journal of Plant Biotechnology
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• v.35 no.4
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• pp.265-274
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• 2008

Glutathione S-transferases (GSTs) are multifunctional proteins encoded by a large gene family divided into Phi, Tau, Theta, Zeta, Lambda and DHAR classes on the basis of sequence identity. The Phi(F) and Tau(U) classes are plant-specific and ubiquitous. Their roles have been defined as herbicide detoxification and responses to biotic and abiotic stresses. Fifty-two members of the GST super-family were identified in the Arabidopsis thaliana genome, 13 members of which belong to the Phi class of GSTs (AtGSTFs). Based on the sequence similarities of AtGSTFs, 11 BAC clones were identified from Brassica rapa. Seven unique sequences of ORFs designated the Phi class candidates of GST derived from B. rapa (BrGSTFs) were detected from these 11 BAC clones by blast search and sequence alignment. Some of BrGSTFs were present in the same BAC clones indicating that BrGSTFs could also be clustered as usual in plant. They were mapped on B. rapa linkage group 2, 3, 9 and 10 and their nucleotide and amino acid sequences were highly similar to those of AtGSTFs. In addition, in silico analysis of BrGSTFs using Korea Brassica Genome Project 24K oligochip and microarray database for cold, salt and drought stresses revealed 15 unigenes to be highly similar to AtGSTFs and six of these were identical to one of BrGSTFs identified in the BAC clones indicating their expression. The sequences of BrGSTFs and unigenes identified in this study will facilitate further studies to apply GST genes to medical and agriculture purposes.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.75-75
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• 2003

Transition of the resting primordial follicle to the growing primary follicle is a critical process for female reproduction, but its mechanism is poorly understood. The present study was conducted to investigate gene expression profile at the primordial-primary follicle transition process. We isolated total RNA of female mouse ovary at day1 (contains only primordial follicles) and day5 (contains primordial and primary follicles) and synthesized cDNA using annealing control primers (ACP; Seegene, Inc., Seoul, Korea). ACP provides annealing specificity and sensitivity to the template and allows to identify only authentic differentially expressed genes (DEGs). We used total 80 ACPs for PCR, observed PCR products on 2% agarose gel, cloned 42 DEGs using TOPO TA cloning vector, sequenced, and analyzed by BLAST search. Sequences of 34 clones significantly matched database entries while 4 clones were novel and 4 clones were EST. Two of 34 genes were specifically expressed only in day 5 ovaries (Sui1-rs1, Apg3p/Aut1p-like), and rest of 32 genes were expressed in both stages but were differential in amount. Differential expression was confirmed using semiquantitative RT-PCR, and there was no false positive. Anx11 and Pepp2-pending were highly expressed genes in day1-, while BPOZ, Ches1, Kcmf1, NHE3, Nid2, Ninj1, SENP3 and Survivin were highly expressed genes in day5-ovary. List of genes would provide insight for further study of mechanism regulating primordial-primary follicle transition.

• The Plant Pathology Journal
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• v.26 no.1
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• pp.8-16
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• 2010

The phytopathogenic fungus Magnaporthe oryzae is a major limiting factor in rice production. To understand the genetic basis of M. oryzae pathogenic development, we previously analyzed a library of T-DNA insertional mutants of M. oryzae, and identified ATMT0879A1 as one of the pathogenicity-defective mutants. Molecular analyses and database searches revealed that a single TDNA insertion in ATMT0879A1 resulted in functional interference with an annotated gene, MGG00056, which encodes a short-chain dehydrogenase/reductase (SDR). The mutant and annotated gene were designated as $MoSDR1^{T-DNA}$ and MoSDR1, respectively. Like other SDR family members, MoSDR1 possesses both a cofactor-binding motif and a catalytic site. The expression pattern of MoSDR1 suggests that the gene is associated with pathogenicity and plays an important role in M. oryzae development. To understand the roles of MoSDR1, the deletion mutant ${\Delta}Mosdr1$ for the gene was obtained via homology-dependent gene replacement. As expected, ${\Delta}Mosdr1$ was nonpathogenic; moreover, the mutant displayed pleiotropic defects in conidiation, conidial germination, appressorium formation, penetration, and growth inside host tissues. These results suggest that MoSDR1 functions as a key metabolic enzyme in the regulation of development and pathogenicity in M. oryzae.