• The korean journal of orthodontics
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• v.33 no.6 s.101
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• pp.485-497
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• 2003

The form and function of the craniofacial structure critically depend on genetic information. With recent advances in the molecular technology, genes that are important for normal growth and morphogenesis of the craniofacial skeleton are being rapidly uncovered, shaping up modem craniofacial biology. One of them is fibroblast growth factor receptor 2 (FGFR2). Specific point mutations in the. FGFR2 gene have been linked to Apert syndrome, which is characterized by premature closure of cranial sutures and craniofacial anomalies as well as limb deformities. To study pathogenic mechanisms underlying craniosynostosis phenotype of Apert syndrome, we used a transgenic approach; an FGFR2 minigene construct containing an Apert mutation (a point mutation that substitute proline at the position 253 to arginine; P253R) was introduced into fertilized mouse germ cells by DNA microinjection. The injected cells were then allowed to develop into transgenic mice. We used a bone-specific promoter (a DNA fragment from the type I collagen gene) to confine the expression of mutant FGFR2 gene to the bone tissue, and asked whether expression of mutant FGFR2 in bone is sufficient to cause the craniosynostosis phenotype in mice. Initial characterization of these mice shows prematurely closed cranial sutures with facial deformities expected from Apert patients. We also demonstrate that the transgene produces mutant FGFR2 protein with increased functional activities. Having this useful mouse model, we now can ask questions regarding the role of FGFR2 in normal and abnormal development of cranial bones and sutures.

• Journal of Life Science
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• v.27 no.8
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• pp.864-872
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• 2017

Equine chorionic gonadotropin (eCG) consists of highly glycosylated ${\alpha}-$ and ${\beta}-subunits$ and is a unique member of the gonadotropin family, because it elicits the response characteristics of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in species other than the horse. To directly assess the biological function of $rec-eCG{\beta}/{\alpha}$, we constructed mammalian expressing vectors of equine luteinizing hormone/chorionic gonadotropin receptors (eLH/CGR). The activity of $rec-eCG{\beta}/{\alpha}$ in vitro assayed in transient transfected CHO-K1 cells and in stably transfected PathHunter Parental cells with eLH/CGR was investigated. $rec-eCG{\beta}/{\alpha}$ was efficiently secreted in the CHO-K1 suspension cell media, and the quantity detected was about 200 mIU/ml from 1 to 7 days after transfection. In the western blot analysis, the $rec-eCG{\beta}/{\alpha}$ protein was broadly identified to be about 40~45 kDa molecular weight. The cAMP stimulation in CHO-K1 cells expressing eLH/CGR was determined to evaluate the activity of $rec-eCG{\beta}/{\alpha}$. The cAMP concentration increased in direct proportion to the concentration of the $rec-eCG{\beta}/{\alpha}$. The $EC_{50}$ value in the transient transfected CHO-K1 cells was $8.1{\pm}6.5ng$. The stable cell lines of eLH/CGR were established in the PathHunter Parental cells expressing ${\beta}-arrestin$. We found that $rec-eCG{\beta}/{\alpha}$ had full LH activity in the PathHunter Parental cells expressing eLH/CGR. The $EC_{50}$ value in transient and stable cells was $5.0{\pm}4.7ng/ml$ and $4.5{\pm}5.2ng/ml$, respectively. These results suggest that $rec-eCG{\beta}/{\alpha}$ has a biological activity in a cell expressing eLH/CGR. These stable cells expressed in PathHunter Parental cells could be useful for elucidating the functional mechanisms of deglycosylated $rec-eCG{\beta}/{\alpha}$ mutants.

• Journal of Animal Science and Technology
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• v.51 no.3
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• pp.185-192
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• 2009

Fatty acid synthase (FASN) is a multi-functional enzyme with a central role in the synthesis of long-chain fatty acid and has been considered as a positional candidate gene for BTA 19 quantitative trait loci (QTL) affecting milk-fat content and fatty acid composition. In this study, we sequenced the FASN gene in several cattle breeds including Hanwoo and imported beef cattle, and identified novel DNA polymorphisms and their linkage relationship in Hanwoo. We found a significant frequency difference of the FASN (AF285607) g.17924 A$\rightarrow$G polymorphism between Hanwoo (70%) and other breeds and this polymorphism has been known for an association with fatty acid composition in Angus. Furthermore, by direct DNA sequencing in 18 unrelated Hanwoo, we identified 27 SNPs including nine novel variations in the FASN gene. Among 27 SNPs identified in the FASN gene, four SNPs were further genotyped in 100 Hanwoo and 96 imported beef cattle, and analyzed for haplotype construction and association with beef quality traits. We performed haplotype block and linkage disequilibrium studies using four selected SNPs. Two different haplotype blocks (block A: g.10568 C$\rightarrow$T and g.11280 G$\rightarrow$ A; block B: g.13125 C$\rightarrow$T and g.17924 G$\rightarrow$A) were constructed and the block A in particular had a very high r2 (0.936), which indicated a nearly complete linkage disequilibrium existed between the g.10568 C$\rightarrow$T and g.11280 G$\rightarrow$A polymorphisms. A total of four major haplotypes (frequency > 0.05) were identified with the four polymorphisms including TATG (0.36), CGCG (0.31), CGTA (0.19) and TACG (0.06). Statistical association analysis revealed that the g.10568 C$\rightarrow$T and g.11280 G$\rightarrow$A polymorphisms in the FASN were significantly associated with meat color (P=0.004) and texture (P=0.0114). The g.13125 C$\rightarrow$T and g.17924 G$\rightarrow$A polymorphisms in the FASN were also significantly associated with back-fat thickness and quantity index (P=0.0179 and 0.0495, respectively). Our findings suggested that the FASN gene polymorphisms may be used for determining the (unsaturated) fatty acid contents and carcass trait in the Hanwoo beef.

• Journal of the Mineralogical Society of Korea
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• v.21 no.3
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• pp.313-320
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• 2008

Hectorite was synthesized under hydrothermal conditions and its physicochemical properties have been investigated in terms of temperature, pH, and organic agent to observe the change of doll basal spacing. The IR, CEC, MB, swelling volume and specific surface area of the hectorite were measured for the characterization. The solid/liquid ratio of hectorite to distilled water before mixing with other materials was also determined for its use as a multi-functional material. The $d_{001}$ basal spacing decreased from $12.63\;\AA$ at room temperature to $10.19\;\AA$ at $650^{\circ}C$ in the heating tests. As the pH of hectorite slurry increased. the $d_{001}$ basal spacing decreased. reaching the lowest value of $13.33\;\AA$ at pH 7 and afterward, increased. All the fool basal spacings of the hectorite increased when it was intercalated with the following solvents: $12.86\;\AA$ in diethyl ether, $13.31\;\AA$ in acetonitrile. $13.59\;\AA$ in methanol, $14.05\;\AA$ in ethanol, $15.69\;\AA$ in acetone, and $17.42\;\AA$ in ethylene glycol. Our IR analysis results were in good agreement with those of other researchers. The CEC, MB, swelling volume and specific surface area of hectorite were determined to be 105 cmol/kg, 80 cmol/kg, $68\sim74ml/2g$ and $213m^{2}/g$, respectively. Also, the hectorite to distilled water ratio of 2 to 100 was found to be most favorable for mixing with other materials such as the solvents mentioned above.

• The Korean Journal of Pharmacology
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• v.30 no.1
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• pp.125-136
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• 1994

To characterize the SR Ca-release channel protein complex of crustacean, $^{45}Ca-release,\;[^3H]ryanodine$ binding, and immunoblot studies were carried out in the crayfish and/or lobster skeletal sarcoplasmic reticulum. Bmax and affinity of crayfish SR to ryanodine were lower than those of lobster SR. AMP (5mM) increased $[^3H]ryanodine$ binding significantly in both vesicles (P<0.05). $Mg^{2+}$(5mM) or tetracaine(1mM) inhibited $[^3H]ryanodine$ binding significantly in both vesicles (P<0.001), but ruthenium red $(10\;{\mu}M)$ inhibited it moderately. In SDS polyacrylamide gel electrophoretic analysis of crayfish SR vesicles, there was a high molecular weight band that showed similar mobility with Ca-release channel protein of lobster skeletal SR, but more rapid mobility (HMWBr) than that of rabbit skeletal SR (HMWBS). Immunoblot analysis showed that polyclonal Ab to lobster skeletal SR Ca-release channel protein was react with HMWBr of crayfish skeletal SR, but not with that of HMWBs of rabbit skeletal SR. ^{45}Ca-release from crayfish skeletal SR vesicles was increased by the increase of extravesicular calcium from $1{\mu}M$ to 1mM. This Ca-release phenomenon was similar, but more sensitive in the low concentration of $Ca^{2+}$, compared to that from lobster SR vesicles. AMP (5mM) or caffeine (10mM) did not affect to $^{45}Ca-release.\;^{45}Ca-release$ was inhibited slightly ($3{\sim}8%\;by\;Mg^{2+}$) (5mM) or tetracaine (1mM), and moderately (23%) by high concentration of ruthenium red $(300\;{\mu}M)$. From the above results, it is suggested that SR Ca-release channel protein of crustacean has different properties from that of the rabbit, and similar properties between crayfish and lobster in functional and immunological aspects, but Ca-release via crayfish channel may be more sensitive to calcium.

• Journal of Applied Biological Chemistry
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• v.57 no.1
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• pp.65-72
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• 2014

Biochar (BC) from biomass pyrolysis is a carbonaceous material that has been used to remove various contaminants in the environment. The eliminatory action for burcucumber (Sicyos angulatus L.) as an invasive plant is being consistently carried out because of its harmfulness and ecosystem disturbance. In this study, burcucumber biomass was converted into BCs at different pyrolysis temperatures of 300 and $700^{\circ}C$ under a limited oxygen condition. Produced BCs were characterized and investigated to ensure its efficiency on antibiotics' removal in water. The adsorption experiment was performed using two different types of antibiotics, tetracycline (TC) and sulfamethazine (SMZ). For the BC pyrolyzed at a high temperature ($700^{\circ}C$), the values of pH, electrical conductivity, and the contents of ash and carbon increased whereas the yield, mobile matter, molar ratios of H/C and O/C, and functional groups decreased. Results showed that the efficiency of BCs on antibiotics' removal increased as pyrolysis temperature increased from 300 to $700^{\circ}C$ (38 to 99% for TC and 6 to 35% for SMZ). The reaction of ${\pi}-{\pi}$ EDA (electron-donor-acceptor) might be involved in antibiotics' adsorption to BCs. BC has potential to be a superior antibiotics' adsorbent with environmental benefit by recycling of waste/invasive biomass.

• Korean Journal of Microbiology
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• v.52 no.4
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• pp.406-414
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• 2016

Among 6 leu codons, CUG is the most frequently used codon in E. coli. It is recognized by leu-tRNA(CAG) encoded by four genes scattered on two chromosomal loci (leuT and leuPQV ). In the process of constructing a strain with no functional leu-tRNA (CAG) gene on chromosome, we made two mutant strains separately, one on leuPQV locus (${\Delta}leuPQV$), and the other on leuT locus [$leuT^*$(GAG)], where the anticodon of leuT was changed from CAG to GAG, thereby altering its recognition codon from CUG to CUC. We attempted to combine these two mutations by transduction using $leuT^*$(GAG) strain as a donor and ${\Delta}leuPQV$ strain as a recipient. Large and small colonies appeared from this transduction. From PCR and DNA sequencing, large colony was confirmed to be the reciprocal recombinant as expected, but the small colonies contained both mutant $leuT^*$(GAG) and wild type leuT (CAG) genes in the cell. This heterozygous diploid strain did not show any unusual morphology under microscopic observation, but, interestingly, it showed a linear growth curve in rich medium with much slower growth rate than wild type cell. It always formed homogenous small colonies in the selection medium, but, when there was no selection, it readily segregated into $leuT^*$(GAG) and leuT (CAG). From these observations, we suggested that the strain with both $leuT^*$(GAG) and leuT (CAG) genes was not a partial diploid (merodiploid), but a full diploid cell having two different chromosomes. We proposed a model explaining how such a heterozygous diploid cell was formed and how and why its growth showed a linear growth curve.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.194-194
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• 2017

Camelina (Camelina sativa L.) is a potential bio-energy crop that has short life cycle about 90 days and contains high amount of unsaturated fatty acid which is adequate to bio-diesel production. Enhancing environmental stress tolerance is a main issue to increase not only crop productivity but also big mass production. CsRCI2s (Rare Cold Inducible 2) are cold and salt stress related protein that localized at plasma membrane (PM) and assume to be membrane potential regulation factor. These proteins can be divide into C-terminal tail (CsRCI2D/E/F/G) or no-tail group (CsRCI2A/B/C/H). However, function of CsRCI2s are less understood. In this study, physiological responses and functional characterization of CsRCI2s of Camelina under salt stress were analyzed. Full-length CsRCI2s (A/B/E/F) and CsPIP2;1 sequences were confirmed from Camelina genome browser. Physiological investigations were carried out using one- or four-week-old Camelina under NaCl stress with dose and time dependent manner. Transcriptional changes of CsRCI2A/B/E/F and CsPIP2;1 were determined using qRT-PCR in one-week-old Camelina seedlings treated with NaCl. Translational changes of CsRCI2E and CsPIP2;1 were confirmed with western-blot using the antibodies. Water transport activity and membrane potential measurement were observed by cRNA injected Xenopus laevis oocyte. As results, root growth rate and physiological parameters such as stomatal conductance, chlorophyll fluorescence, and electrolyte leakage showed significant inhibition in 100 and 150 mM NaCl. Transcriptional level of CsPIP2;1 did not changed but CsRCI2s were significantly increased by NaCl concentration, however, no-tail type CsRCI2A and CsRCI2B increased earlier than tail type CsRCI2E and CsRCI2F. Translational changes of CsPIP2;1 was constitutively maintained under NaCl stress. But, accumulation of CsRCI2E significantly increased by NaCl stress. CsPIP2;1 and CsRCI2A/B/E/F co-expressed Xenopus laevis oocyte showed decreased water transport activity as 61.84, 60.30, 62.91 and 76.51 % at CsRCI2A, CsRCI2B, CsRCI2E and CsRCI2F co-expression when compare with single expression of CsPIP2;1, respectively. Moreover, oocyte membrane potential was significantly hyperpolarized by co-expression of CsRCI2s. However, higher hyperpolarized level was observed in tail-type CsRCI2E and CsRCI2F than others, especially, CsRCI2E showed highest level. It means transport of $Na^+$ ion into cell is negatively regulated by expression of CsRCI2s, and, function of C-terminal tail is might be related with $Na^+$ ion influx. In conclusion, accumulation of NaCl-induced CsRCI2 proteins are related with $Na^+$ ion exclusion and prevent water loss by CsPIP2;1 under NaCl stress.

• Journal of the Korean Society of Food Science and Nutrition
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• v.14 no.4
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• pp.345-352
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• 1985

To characterize the protein from arrowroot leaf, proteins were extracted and separated from arrowroot leaf, then its amino acid composition and functional properties were studied. Protein in arrowroot leaf was consisted of 18.5% albumin, 33.5% globulin, 34.0% glutelin, 6.2% protamine and 7.8% insoluble residues. The rates of precipitation of proteins which extracted with water, 1M NaCl, and 0.015N NaOH as a solvent were 84.7% (at pH 3.0), 76.4% (at pH 2.5) and 86.4% (at pH 4.0), respectively. The extracted proteins were separated up to about 90% by organic solvents such as ethanol and acetone at 80% concentration, Composition of arrowroot leaf protein concentrates were: $1{\sim}2%$ moisture, $59{\sim}67%$ protein, $4{\sim}8%$ ash and $5{\sim}6%$ (dialyzed concentrates) or $1{\sim}2%$ (acetone-treated ones) lipid. Main amino acids of the concentrates were aspartic acid, glutamic acid and glycine. Solubility profile of the concentrates according to pH was typical. The minimum solubility (below pH 5.0) of acetone extracted protein concentrates was lower than that of unextracted ones, whereas the reverse was true for pH value above this region. Bulk density, water and fat absorption of the concentrates were attributable to correlation to the treatment of acetone. And the bulk density of the concentrates was negatively correlative to both water and fat absorption. Emulsifying and foaming properties were not varied with the treatment of acetone.

• Korean Journal of Environmental Agriculture
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• v.35 no.1
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• pp.79-86
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• 2016

BACKGROUND: This study was initiated to quantitatively evaluate the effects of five heavy metals on the growth and P removal efficiencies of Alcaligenes sp., known as the Phosphorus Accumulating Organisms (PAOs). It was cultivated in the batch system with five heavy metals, such as Cd, Cu, Zn, Pb and Ni, added in single and binary mixtures, respectively.METHODS AND RESULTS: IC₅₀ (half of inhibition concentration of bacterial growth) and EC₅₀ (half of effective concentration of phosphorus removal Efficiencies) were used to quantitatively evaluate the effects of heavy metals on the growth and phosphorus removal Efficiencies of Alcaligenes sp. In addition, Additive Index Value (A.I.V.) method was used to evaluate the interactive effects between Alcaligenes sp. and heavy metals. As a result, as the five heavy metals were singly added to Alcaligenes sp., the greatest inhibitory effects on the growth and P removal efficiencies of each bacteria was observed in the cadmium (Cd). In the binary mixture treatments of heavy metals, the treatments of lowest IC₅₀ and EC₅₀ were the Cd + Cu treatment. Based on the IC₅₀ and EC₅₀ of the binary mixtures of heavy metals treatments, most interactive effects between the heavy metals were found to be antagonistic.CONCLUSION: Based on the results obtained from this study, it appears that they could provide the basic information about the toxic effects of the respective treatments of single and binary mixtures of heavy metals on the growth and P removal efficiencies of Alcaligenes sp. through further study about the characterization of functional proteins involved in toxic effects of heavy metals.