• Journal of Plant Biotechnology
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• v.41 no.3
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• pp.107-115
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• 2014

Vegetable oils (triacylglycerols) produced mainly in seeds of plants are used for valuable foods that supply essential fatty acids for humans as well as industrial raw materials and biofuel production. As the demanding for vegetable oils has increased, plant metabolic engineering to produce triacylglycerols in biomass such as leaves has been considered and explored for alternative source of vegetable oils. Leaves are genetically programmed to supply the fixed carbon by photosynthesis to other organs for plant development and growth. Therefore, in order to produce and accumulate triacylglycerols in leaves, one should take account of multiple metabolic pathways such as carbon flux, competition of carbohydrate and fatty acid biosynthesis, and triacylglycerols turnover in leaves. The recent metabolic engineering strategy has showed potential in which the co-expression of three genes WRINKLED1, DGAT1, and OLEOSIN involved in the critical step for increasing the fatty acid synthesis, accumulating triacylglycerols, and protecting triacylglycerols, respectively produced higher amount of vegetable oils in leaves. Developing of genetically engineered plants producing vegetable oil in biomass at non-agricultural lands will be promising to the future success of the field.

• Journal of Plant Biotechnology
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• v.37 no.2
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• pp.220-227
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• 2010

Fatty acids in seed oil from plants are essential for human nutrients and have been used for industrial purpose. The growing demands of seed oil as food resources and feedstocks for industrial uses have attempted to modify fatty acid composition and to increase oil content in transgenic plants. However, production of unusual fatty acids in transgenic plants are limited, which is not synthesized the level same as original plants. This bottleneck was common for production of several unusual fatty acids in transgenic plants and suggests that there is different for substrate preference in oil metabolic pathway enzymes between host oil plants and original wild plants. Review of acyltransferases involved in acyl-editing and seed oil accumulation of oil plant and wild-plant producing unusual fatty acids will design strategies to maximize the production of unusual fatty acids in transgenic plants. In here, we identified eleven acyltransferase genes in castor based on sequence homology, which will be useful to increase hydroxy unusual fatty acids in transgenic plants.

• KSBB Journal
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• v.17 no.4
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• pp.370-375
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• 2002

Isolation of a gene and determination of its expression pattern are essential in understanding its function. Among the genes localized in 12ql3, stSG3435 EST was chosen to study its expression pattern. The full-length CDNA was cloned by screening of human brain CDNA library and its sequence was determined by serial deletion followed by automated sequencing of the clones with overlapping fragments. The sequence analysis revealed that stSG 3435 CDNA displayed 100% identity to human MYGI and 86% identity to mouse melanocyte proliferation gene-1 (Gamm 1) originally identified from melanocyte, suggesting that MYGI determined by Northern blot analysis revealed the strongest expression in testes with ubiquitous expression in all the tissues tested. In order to investigate the cellular localization of its protein product, the green fluorescence protein gene was fused into the full-length coding sequence of MYGI, Transfection of the fusion construct followed by confocal microscopy resulted in the green fluorescence signal as a punctate state in cytoplasm indication that MYGI was localized in one of the cellular organelles.

• Genomics & Informatics
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• v.9 no.4
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• pp.152-160
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• 2011

Hypertension is the major factor of most death and high blood pressure (BP) can lead to stroke, myocardial infarction and cardiac failure. Moreover, hypertension is strongly correlated with body mass index (BMI). Although the exact causes of hypertension are still unclear, some of genetic loci were discovered from genome-wide association study (GWAS). Therefore, it is essential to study genetic variation for finding more genetic factor affecting hypertension. The purpose of our study is to conduct a CNV association study for hypertension-related traits, BP and BMI, in Korean individuals. We identified 2,206 CNV regions from 3,274 community-based Korean participants using the Affymetrix Genome-Wide Human SNP Array 6.0 platform and performed a logistic regression analysis of CNVs with two hypertension-related traits, BP and BMI. Moreover, the 4,692 participants in an independent cohort were selected for respective replication analyses. GWAS of CNV identified two loci encompassing previously known hypertension-related genes: LPA (lipoprotein) on 6q26, and JAK2 (Janus kinase 2) on 9p24, with suggestive p-values (0.0334 for LPA and 0.0305 for JAK2 ). These two positive findings, however, were not evaluated in the replication stage. Our result confirmed the conclusion of CNV study from the WTCCC suggesting weak association with common diseases. This is the first study of CNV association study with BP and BMI in Korean population and it provides a state of CNV association study with common human diseases using SNP array.

• Journal of dental hygiene science
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• v.13 no.3
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• pp.321-329
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• 2013

Although it has been reported that lysyl oxidase (LOX) is involved in odontoblastic differentiation, the role of LOX on odontoblastic differentiation by hydrogen peroxide ($H_2O_2$) have not been clarified. In the present study, we investigated whether $H_2O_2$, reactive oxygen species (ROS), is modulated the messenger RNA (mRNA) expression and activity of LOX during odontoblastic differentiation of human dental pulp (HDP) cells. The mRNA expression was quantified by reverse transcriptase polymerase chain reaction (RT-PCR) analysis, and LOX enzyme activity was measured by high sensitive fluorescent assay. Expression of the odontoblastic differentiation marker genes were assessed in the presence and absence of specific small interfering RNAs (siRNAs) of the LOX and LOXL. The $H_2O_2$-induced mRNA expression of LOX family was significant reduction of LOX, LOXL, and LOXL3 mRNA levels in HDP cells. LOX enzyme activity was increased at $H_2O_2$ 0.3 mM for 24 hours. The mRNA expression of alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OCN) was inhibited by LOX- and LOXL-specific siRNAs whereas the mRNA expression of dentin matrix protein1 (DMP1), and dentin sialophosphoprotein (DSPP) was inhibited by LOX-specific siRNA. In LOX enzyme activity, siRNA-induced knockdown of both LOX and LOXL inhibited the total amine oxidase activity in HDP cells, as in the case of mRNA expression. In conclusion, the essential role of $H_2O_2$ on odontoblastic differentiation suggests that its regulation by LOX may have pharmacologic importance in HDP cells.

• Journal of Microbiology and Biotechnology
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• v.28 no.6
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• pp.997-1006
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• 2018

As shown during the 2009 pandemic H1N1 (A(H1N1)pdm09) outbreak, egg-based influenza vaccine production technology is insufficient to meet global demands during an influenza pandemic. Therefore, there is a need to adapt cell culture-derived vaccine technology using suspended cell lines for more rapid and larger-scale vaccine production. In this study, we attempted to generate a high-growth influenza vaccine strain in MDCK cells using an A/Puerto/8/1934 (H1N1) vaccine seed strain. Following 48 serial passages with four rounds of virus plaque purification in MDCK cells, we were able to select several MDCK-adapted plaques that could grow over $10^8PFU/ml$. Genetic characterization revealed that these viruses mainly had amino acid substitutions in internal genes and exhibited higher polymerase activities. By using a series of Rg viruses, we demonstrated the essential residues of each gene and identified a set of high-growth strains in MDCK cells ($PB1_{D153N}$, $M1_{A137T}$, and $NS1_{N176S}$). In addition, we confirmed that in the context of the high-growth A/PR/8/34 backbone, A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2), and A/environment/Korea/deltaW150/2006 (H5N1) also showed significantly enhanced growth properties (more than $10^7PFU/ml$) in both attached- and suspended-MDCK cells compared with each representative virus and the original PR8 vaccine strain. Taken together, this study demonstrates the feasibility of a cell culture-derived approach to produce seed viruses for influenza vaccines that are cheap and can be grown promptly and vigorously as a substitute for egg-based vaccines. Thus, our results suggest that MDCK cell-based vaccine production is a feasible option for producing large-scale vaccines in case of pandemic outbreaks.

• International Journal of Oral Biology
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• v.36 no.4
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• pp.173-178
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• 2011

Tumor necrosis factor alpha ($TNF{\alpha}$) is a multifunctional cytokine that is elevated in inflammatory diseases such as atherosclerosis, diabetes and rheumatoid arthritis. Recent evidence has suggested that ${\beta}2$ adrenergic receptor (${\beta}2AR$) activation in osteoblasts suppresses osteogenic activity. In the present study, we explored whether $TNF{\alpha}$ modulates ${\beta}AR$ expression in osteoblastic cells and whether this regulation is associated with the inhibition of osteoblast differentiation by $TNF{\alpha}$. In the experiments, we used C2C12 cells, MC3T3-E1 cells and primary cultured mouse bone marrow stromal cells. Among the three subtypes of ${\beta}AR$, ${\beta}2$ and ${\beta}3AR$ were found in our analysis to be upregulated by $TNF{\alpha}$. Moreover, isoproterenol-induced cAMP production was observed to be significantly enhanced in $TNF{\alpha}$-primed C2C12 cells, indicating that $TNF{\alpha}$ enhances ${\beta}2AR$ signaling in osteoblasts. $TNF{\alpha}$ was further found in C2C12 cells to suppress bone morphogenetic protein 2-induced alkaline phosphatase (ALP) activity and the expression of osteogenic marker genes including Runx2, ALP and osteocalcin. Propranolol, a ${\beta}2AR$ antagonist, attenuated this $TNF{\alpha}$ suppression of osteogenic differentiation. $TNF{\alpha}$ increased the expression of receptor activator of NF-${\kappa}B$ ligand (RANKL), an essential osteoclastogenic factor, in C2C12 cells which was again blocked by propranolol. In summary, our data show that $TNF{\alpha}$ increases ${\beta}2AR$ expression in osteoblasts and that a blockade of ${\beta}2AR$ attenuates the suppression of osteogenic differentiation and stimulation of RANKL expression by $TNF{\alpha}$. These findings imply that a crosstalk between $TNF{\alpha}$ and ${\beta}2AR$ signaling pathways might occur in osteoblasts to modulate their function.

• Journal of Life Science
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• v.16 no.7 s.80
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• pp.1112-1118
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• 2006

Bordetella bronchiseptica, the agent of swine atrophic rhinitis and kennel cough in dogs, is a mucosal pathogen and produces the hydroxamate type alcaligin siderophore under iron-limited conditions. Genes involved in alcaligin siderophore biosynthesis are contained in an alcABCDE operon. In order to provide direct evidence for the role of AlcA in alcaligin biosynthesis, we needed a B. bronchiseptica mutant carrying alcA gene deletion. A 0.6 kb alcA 5'-flanking and 0.7kb 3'-flanking DNA fragments were PCR amplified with the use of pCP1.11 as a template DNA. The 5'-and 3'-flanking DNA fragments were joined in a suicide plasmid, resulting in a recombinant suicide plasmid pDM1. After introduction of pDM1 into B. bronchiseptica by conjugation, the allelic exchange technique was performed and a B. bronchiseptica alcA deletion mutant, named B. bronchiseptica H1, was obtained. The mutant strain produced reduced amount of siderophore as expected. When a plasmid containing complete alcA gene was transformed back into the mutant, the complemented mutant recovered ability of siderophore production. These results indicated that AlcA is one of essential components for the alcaligin siderophore biosynthesis. The mutant strains obtained in this study will be used in the further studies for the biochemical function of AlcA.

• Journal of Life Science
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• v.22 no.12
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• pp.1600-1607
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• 2012

The matrix metalloproteinase (MMP) family plays essential roles in physiological processes such as embryonic development, angiogenesis, wound healing, and tissue homeostasis as a consequence of MMPr capacity for breaking down many types of extracellular matrix proteins. Imbalanced regulation of MMP expression can also lead to pathological conditions such as tumor progression. We recently reported that the Drosophila Mmp1 gene is highly expressed in the digestive tract and is required for the maintenance of intestinal homeostasis such as by restriction of uncontrolled intestinal stem cell proliferation. However, the regulatory mechanisms of MMP gene expression in the intestine remain unclear. In this study, we determined that the expression of Mmp1 is regulated by the homeodomain transcription factor Caudal. Experiments using the targeted expression of Caudal under the regulation of Gal4-UAS system indicated that endogenous Caudal is required for the Mmp1 gene expression in the adult Drosophila intestine and that exogenous Caudal induces Mmp1 expression. Transient transfection experiments indicated that Caudal can activate the promoter activity of Mmp1 and that several putative Caudal binding sites in the 5'-flanking region of the Mmp1 gene may be critical to the upregulation by Caudal. Our data suggest that Mmp1 is one of the target genes of Caudal in physiological normal condition and in tumorigenesis.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.3
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• pp.166-172
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• 2006

Metallo-${\beta}$-lactamase (MBL) can hydrolyze all ${\beta}$-lactams except monobactams and frequently coexists with various antibiotic resistance genes such as aminoglycoside resistance, sulfonamide resistance gene, etc. Therefore, the effective antibiotics against infections by these bacteria are markedly limited or can't even be found. We tried to search in-vitro antimicrobial combinations with synergistic effects for a VIM-2 type MBL producing Pseudomonas aeruginosa, isolated from clinical specimen. On the selection of antibiotic combinations with synergistic effects, we performed a one disk synergy test, modified Pestel's method, in agar without aztreonam (AZT). The bacteriostatic synergistic effects of this tests were scored as $S_1$ (by susceptibility pattern in agar without antibiotics), $S_2$ (by the change of susceptibility in agar with or without antibiotics) and $S_3$ ($S_1$ + $S_2$) and was classified into weak (1 point), moderate (2 points) and strong (3 points) by $S_3$ score. Subsequently, we carried out the time-killing curve for the antibiotic combinations with the strong synergistic bacteriostatic effect. One VIM-2 type MBL producing P. aeruginosa confirmed by the PCR showed all resistance against all ${\beta}$-lactams except AZT, aminoglycoside and ciprofloxacin. In the one disk synergy test, this isolate showed a strong bacteriostatic synergistic effect for the antibiotic combination of AZT and piperacillin-tazobactam (PIP-TZP) or AZT and amikacin (AN). On the time-killing curve after six hours of incubation, the colony forming units (CFUs/mL) of this bacteria in the medium broth with both combination antibiotics were decreased to 1/18.7, 1/17.1 of the least CFUs of each single antibiotics. The triple antibiotic combination therapy including AZT, PIP-TZP and AN was shown to be significantly synergistic after 8 hrs of exposure. In a VIM-2 MBL producing P. aeruginosa with susceptibility for AZT, the triple antibiotic combination therapy including AZT, PIP-TZP and AN may be considered as an alternative antibiotics modality against the infection by some MBL type. But the antimicrobial combination therapy for many more MBL producing isolates is essential to know as soon as possible for the selection of effective treatment against the infection by this bacteria.