• Journal of Microbiology and Biotechnology
• /
• v.16 no.6
• /
• pp.933-938
• /
• 2006

Decaprenyl diphosphate synthase (DPS) is the key enzyme for the production of coenzyme $Q_{10}$ ($CoQ_{10}$). A dps gene from Sinorhizobium meliioti KCCM 11232 (IFO 14782) was isolated by PCR and then cloned in Escherichia coli. DNA sequencing analysis revealed an open reading frame of 1,017 bp encoding a 338-amino-acid protein. The protein was identical at the 98% level to the putative octaprenyl diphosphate synthase (IspB) of S. meliloti 1021. The deduced amino acid sequence included the DDxxD domains conserved in the majority of the prenyl diphosphate synthases. Heterologous expression in E. coli BL21 (DE3) was carried out, and the $CoQ_{10}$ produced was then analyzed by HPLC. E. coli BL21 (DE3) harboring the dps gene from S. melioti produced CoQ$_{10}$ in addition to endogenous coenzyme Q$_8$ (CoQ$_8$), whereas wild-type E. coli BL21 (DE3) host did not have the ability of producing CoQ$_{10}$. The results suggest that the putative dps from S. meliloti KCTC 2353 encoded the DPS.

• Molecules and Cells
• /
• v.43 no.6
• /
• pp.581-589
• /
• 2020

Neurons have multiple dendrites and single axon. This neuronal polarity is gradually established during early processes of neuronal differentiation: generation of multiple neurites (stages 1-2); differentiation (stage 3) and maturation (stages 4-5) of an axon and dendrites. In this study, we demonstrated that the neuron-specific n-glycosylated protein NELL2 is important for neuronal polarization and axon growth using cultured rat embryonic hippocampal neurons. Endogenous NELL2 expression was gradually increased in parallel with the progression of developmental stages of hippocampal neurons, and overexpression of NELL2 stimulated neuronal polarization and axon growth. In line with these results, knockdown of NELL2 expression resulted in deterioration of neuronal development, including inhibition of neuronal development progression, decreased axon growth and increased axon branching. Inhibitor against extracellular signal-regulated kinase (ERK) dramatically inhibited NELL2-induced progression of neuronal development and axon growth. These results suggest that NELL2 is an important regulator for the morphological development for neuronal polarization and axon growth.

• KSCE Journal of Civil and Environmental Engineering Research
• /
• v.35 no.6
• /
• pp.1347-1356
• /
• 2015

Since 1970 transportation investment has been made rapidly, however the doubt on duplication and overinvestment of transportation facility also has been brought up. Thus the necessity for reasonable estimation of investment scale increases. In this paper various researches related to this subject are reviewed and categorized according to their methodologies, namely, the methodology using international comparison, trend of investment policy and endogenous economy development model. Based on these methodologies, AHP is applied to calculate proper investment scale. The result shows that the budget for satisfying the investment scale is estimated about 19.51~24.49 trillion won. This value is 4.28~9.26 trillion won higher than the proper investment budget established in National Fiscal Management Plan. In the future, it is necessary to establish systematic and efficient transportation investment plan by developing transportation policy model which can assess the economic feasibility, social equity and environmental soundness etc. at the same time.

• Journal of Environmental Science International
• /
• v.20 no.8
• /
• pp.943-951
• /
• 2011

We investigated the effects of temperature changes on the oxygen consumption rhythm in Japanese eels, Anguilla japonica, using an automatic intermittent flow respirometer (AIFR). The endogenous rhythm of the oxygen consumption rate (OCR) in the eels (n = 18; 44-74 cm, 145-690 g), freshly collected by bag net from estuaries, was nearly synchronous with the tidal pattern of the estuarine collection site. The magnitude of mean OCR (mOCR) of eels showed variable range of 82.2 - 116.5 ml $O_2\;kg^{-1}ww \;h^{-1}$ under constant conditions. In case of increasing temperature from 25 to $38^{\circ}C$, the OCR of eels exhibited a gradually increasing trend with a rhythmic pattern until $36^{\circ}C$. Above $36^{\circ}C$ the rhythms of the OCR dampened and the OCR decreased rapidly at around $36-37^{\circ}C$. The OCR of the eels exhibited the maximum value at $38^{\circ}C$, and then it sharply decreased. The results suggested that the critical thermal maximum (CTM) regarding the endogenous rhythms of the eels was at around $36-37^{\circ}C$ when water temperature increased at $0.5^{\circ}C$/14 h following the acclimation at $25^{\circ}C$. In case of decreasing temperature ($0.5^{\circ}C$/14 h) from 25 to $0^{\circ}C$, the OCR of the eels displayed a abrupt decrease up to $23^{\circ}C$, and between at 23 and $20^{\circ}C$, there was an agitation which showed a slight increase in the OCR with a duration of 1-2 days. Below $9^{\circ}C$, the OCR rhythm of the eels showed a constant state regardless of temperature decreasing. These results suggest that the Japanese eel has an upper incipient lethal temperature at $36^{\circ}C$, with a lower thermal limit at $9^{\circ}C$. The biochemical aspects of the eels influenced by water temperature need to be further studied.

• Journal of Aquaculture
• /
• v.9 no.4
• /
• pp.445-452
• /
• 1996

Endogeneous activities of ${\beta}-galactosidase$-like enzyme in various tissues from several finfishes and shellfishes were examined by histochemical analysis based on X-gal staining and by fluorimetric measurement using 4-methylumbelliferyl-${\beta}$-D-galactoside (4-MUG). Species used in this study were 3 freshwater fishes, mud loach (Misgurnus mizolepis), common carp (Cyprinus carpio) and tilapia (Oreochromis niloticus) ; 3 marine fishes, olive flounder (Paralichthys olivaceus), stone flounder (Kareius bicoloratus) and marbled sole (Limanda yokohamae) ; and 4 shellfishes, abalone (Haliotis discus hannai), Pacific oyster (Crassoskra gigas), pearl oyster (Pinctada fucata martensii) and ark shell (Anadara broughtonii). The activities of ${\beta}-galactosidase$-like enzyme in all finfishes examined were significantly different among species, with the wide variations between tissues in a species. In general, the tissues such as kidney, intestine and liver were ones which showed the significantly higher values in 4-MUG fluorimetry and deeper staining patterns in X-gal analysis compared to other tissues. On the other hand, serum and muscle revealed the significantly lower activities than others did, regardless of species. Shellfishes were also found to have endogenous activities of ${\beta}-galactosidase$-like enzyme which were significantly varied depending on both species and organs in a species. Hepatopancreas from all shellfishes examined showed the deepest pattern in X-gal staining and also the highest value in 4-MUG analysis, while activities of ${\beta}-galactosidase$-like enzyme in adductor muscles and mantle muscles from all shellfish species in this study except Pacific oyster were negligible : Pacific oyster had the significant activity of this enzyme in muscle tissues. Putative endogenous lacZ fragment was amplified from both finfishes and shellfishes by polymerase chain reaction (PCR). The molecular size of PCR products was about 510 bp, and there was no difference in size among species examined.

• Horticultural Science & Technology
• /
• v.28 no.5
• /
• pp.750-756
• /
• 2010

This study was performed to investigate the role of phytohormones in the bulbing of garlic in order to assess the yield and quality. The effect on endogenous plant hormones such as gibberellin (GA) content was also examined during growth stage i.e. clove differentiation to bulbing in garlic. More than 18 gibberellins in garlic were identified with extensive gas chromatograph-mass spectrometry-selected ion monitoring (GC-MS-SIM) quantitative analysis. The results showed that GAs were biosynthesized by both non C-13 hydroxylation pathway (NCH) and early C-13 hydroxylation pathway (ECH) in garlic plant. It was also revealed that NCH pathway leading to synthesis of bioactive $GA_4$ was the more prominent GA biosynthesis pathway than ECH pathway in which bioactive $GA_1$ was synthesized. Total GAs level was gradually increased from clove differentiation to bulbing and later decreased, which portrays the active role of GA in differentiation. The biosynthesis ratio of bioactive $GA_4$ and $GA_1$ concentration was similar to that of total GAs content, which was closely related with bulb development in garlic.

• Journal of Agricultural Extension & Community Development
• /
• v.21 no.2
• /
• pp.1-28
• /
• 2014

Private consulting firms that are currently participating in rural area development projects with a bottom-up approach are involved in nearly all areas of rural area development, and the policy environment that emphasizes the bottom-up approach will further expand their participation. Reviews of private consulting firms, which started out with high expectations in the beginning, are now becoming rather negative. Expertise is the key issue in the controversy over private consulting firms, and the analysis tends to limit the causes of the problems within firms. This study was conducted on the premise that the fixation on cause and structure results in policy issues in the promotion process. That is because the government authorities are responsible for managing and supervising the implementation of policies, not developing the policies. The current issues with consulting firms emerged because of the hasty implementation of private consulting through the government policy trend without sufficient consideration, as well as the policy environment that demanded short-term outcomes even though the purpose of bottom-up rural area development lies in the ideology of endogenous development focused on the changes in residents' perceptions. Research was conducted to determine how the problems of private consulting firms that emerged and were addressed in this context influenced the consulting market, using current data and based on the firms' business performance. In analyzing the types, firms were divided into three groups: top performers including market leaders (9), excellent performers (36), and average performers (34). An analysis of the correlation between the business performance of each type and managerial resources such as each firm's expertise revealed that there was only a correlation between human resources and regional development in excellent performers, and none was found with the other types. These results imply that external factors other than a firm's capabilities (e.g., expertise) play a significant role in the standards of selecting private consulting firms. Thus, government authorities must reflect on their error of hastily adopting private consulting firms without sufficient consideration and must urgently establish response measures.

• Journal of Plant Biotechnology
• /
• v.40 no.1
• /
• pp.18-26
• /
• 2013

Genetically modified (GM) rice Agb0101, which expresses the insecticidal toxin modified cry1Ac (mcry1Ac1) gene, was developed by the Rural Development Administration in Korea. To monitor the probable release of Agb0101 in the future, it is necessary to develop a reliable detection method. Here, we developed the PCR detection method for monitoring and tracing of GM rice. The primer pair (RBEgh-1/-2) from a starch branching enzyme (RBE4) gene was designed as an endogenous reference, giving rise to an expected PCR amplicon of 101 bp. For the qualitative PCR detection, construct- and event-specific primers were designed on the basis of integration sequence of T-DNA. Event-specific PCRs amplified specifically 5'- or 3'-junction region spanning the native genome DNA and the integrated gene construct, while none of amplified product was shown on crops, rice varieties, and other insect-resistant transgenic rice lines. The event-specific real-time PCR method was performed using TaqMan probe and plasmid pRBECrR containing both rice endogenous gene RBE4 sequence and 5'-junction sequence as the reference molecule. The absolute limit of quantification (LOQ) of real-time PCR was established with around 10 copies for one plasmid molecule pRBECrR. Thereafter, the different amounts of transgenic rice (1, 3, 5, and 10%, respectively) were quantified by using the established real-time PCR method, with a range below 19.55% of the accuracy expressed as bias, 0.06-0.40 of standard deviation (SD) and 3.80-7.01% of relative standard deviations (RSD), respectively. These results indicate that the qualitative and quantitative PCR methods could be used effectively to detect the event Agb0101 in monitoring and traceability.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 2001.11a
• /
• pp.108-108
• /
• 2001

Axin2/Conductin/Axil and its ortholog Axin are negative regulators of the Wnt signaling pathway, which promote the phosphorylation and degradation of ${\beta}$-catenin. While Axin is expressed ubiquitously, Axin2 mRNA was seen in a restricted pattern during mouse embryogenesis and organogenesis. Because many sites of Axin2 expression overlapped with those of several Wnt genes, we tested whether Axin2 was induced by Wnt signaling. Endogenous Axin2 mRNA and protein expression could be rapidly induced by activation of the Wnt pathway, and Axin2 reporter constructs, containing a 5.6 kb DNA fragment including the promoter and first intron, were also induced. This genomic region contains eight Tcf/LEF consensus binding sites, five of which are located within longer, highly conserved non-coding sequences. The mutation or deletion of these Tcf/LEF sites greatly diminished induction by ${\beta}$-catenin, and mutation of the Tcf/LEF site T2 abolished protein binding in an electrophoretic mobility-shift assay. These results strongly suggest that Axin2 is a direct target of the Wnt pathway, mediated through Tcf/LEF factors. The 5.6 kb genomic sequence was sufficient to direct the tissue specific expression of d2EGFP in transgenic embryos, consistent with a role for the Tcf/LEF sites and surrounding conserved sequences in the in vivo expression pattern of Axin2. Our results suggest that Axin2 participates in a negative feedback loop, which could serve to limit the duration or intensity of a Wnt-initiated signal.

• Clinical and Experimental Reproductive Medicine
• /
• v.26 no.2
• /
• pp.219-223
• /
• 1999

Ovarian development of the vitrified neonatal ovaries after orthotopical transplantation into the ovariectomized adult recipient mouse were observed. Ovaries were collected from the neonatal females on day of birth and grouped for fresh, vitrification for 1-minute, and 3-minute. Vitrified and thawed neonatal ovaries were orthotopically transplanted into ovarian bursa of the adult mice from which endogenous ovaries have removed just prior to the transplantation (1 minute: n=25; 3 minutes n=23). Fresh ovarian tissue transplanted (n=25) mice were included as control groups. Returning of the estrus cycles and the survival and development of the transplanted ovaries were evaluated. Intact ovaries from neonatal, and four weeks old mice were used for comparison of the ovarian development as in vivo-developed control. From 2 weeks after transplantation, 64%, 36%, and 75% of the transplanted mice showed return of the estrus cycles in fresh, 1-minute, and 3-minute groups, respectively. Four weeks after transplantation, all mice were sacrificed and ovarian tissues were recovered for histological analysis. 57.1%, 33.3%, and 64.7% mice in fresh, 1-minute, and 3-minute groups, respectively, had survived ovaries with follicles at various stages of growth from primordial to preovulatory follicles. Corpus lutea were also observed. Results of the present study suggest that 1) normal folliculogenesis has initiated in vivo after vitrification, and 2) the vitrification may be used as a preservation method for ovarian tissues for establishment of ovarian tissue bank.