• BMB Reports
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• v.29 no.1
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• pp.88-91
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• 1996

The anaerobically expressed gene aeg-46.5, which had been identified by the operon fusion technique with a hybrid bacteriophage of ${\lambda}$ and Mu, ${\lambda}$placMu53, was studied for its expression pattern and growth. The expression of aeg-46.5 was studied in the wild-type cell and mutant cells that have mutation (s) in the control gene of anaerobic respiration (fnr) and nitrate response (narL and narP). The ${\beta}$-galactosidase reporter gene showed maximum expression in narL host after two hours of aerobic to anaerobic switch in M9-Glc-nitrate medium. Both 40 mM and 100 mM concentrations of nitrate ion in the medium had little effect on expression level. We propose that aeg-46.5 is subject to multiple regulations of anaerobic activation by Fnr, nitrate activation by NarP and repression mediated by NarL.

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.436-442
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• 1999

The aeg-46.5 operon of Escherichia coli is induced by nitrate and anaerobic conditions. Positive regulators Fnr and NarP, and a negative regulator NarL control the expression of the aeg-46.5. It has two symmetry regions [6], one of which is located between +37 and +56 bp from the 5'end of the anaerobic transcription initiation site. In this study, mutagenized symmetry regions were transferred from plasmid to chromosome by homologous recombination to evaluate the mutation as a single copy in the fnr, narL, narP, and narL-narP double mutant background. The expressions of the aeg-46.5 operon with these mutations indicated that the control was not through the possible stem-loop structure. Whether there is a protein that mediates this control remains to be seen. The results from the narL-narP double mutant indicated that the anaerobic Fill induction was independent of NarL repression.

• Nutrition Research and Practice
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• v.5 no.1
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• pp.66-72
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• 2011

The purpose of this study was to assess the metabolic profile and nutrient intake data in Korean adolescents according to exercise regularity using the forth Korean National Health and Nutrition Examination Survey from 2008 data. A total of 834 Korean adolescents (440 boys, 394 girls) aged 12 to 18 years were classified into exercising male (EM), non-exercising male (NM), exercising female (EF), and non-exercising female (NF) groups. EM exhibited significantly higher weight (P<0.001), waist circumference (WC) (P<0.01) and body mass index (EMI) (P<0.001) than NM. EF showed significantly greater height (P<0.01), weight (P<0.001), we (P<0.001), and BMI (P<0.001) than NF. Although ratios of macronutrient intake were within the appropriate range in all groups, energy, riboflavin, vitamin e and calcium were assessed as in adequate by the nutritional adequacy ratio (NAR) in all groups. Moreover, EF and NF had inadequate intake of vitamin A and iron according to NAR, respectively. There were significant correlations between height and NAR protein (r=0.249, P<0.001), and mean adequacy ratio (MAR) (r=0.177, P<0.01), and between weight and NAR protein (r=0.180, P<0.01), and MAR(r=0.136, P<0.05) in EM. On the other hand, there were significant correlations between weight and NAR protein (r=0.270, P<0.01), we and NAR protein (r=0.271, P<0.01), and BMI and NAR protein (r=0.326, P<0.01) in NM. There were significant correlations height and NAR Fe (r=0.153, P<0.05) in EF. However, there were no significant correlations between metabolic factors and nutritional adequacy in NF. Although we noted no significant differences in MAR between the groups, the exercising groups showed higher MAR values than the non-exercising groups. Therefore, practicing of ideal dietary behaviors appears to be induced through physical activity and regular exercise in Korean adolescents.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.762-766
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• 2001

The nar promoter of Escherichia coli was known to induce maximally under anaerobic or microaerobic conditions in the presence of nitrate. In this study, the nar promoter was tested to see whether the expression level of a reporter gene which fused lacZ gene at nar promoter's downstream, in the some gram negative host strains(Agrobacterium, Pseudomonas and Rhizobium). A nar promoter system(Combination of nar promoter and gram negative strain) was grown under aerobic conditions to absorbance at 600 nm of nearly 2.0 and then, the nar promoter was induced by lowering DO to 1-2% with alternating microaerobic and aerobic condition in the fermentor cultures, using different gram negative hosts. For a wild type nar promoter (pNW61), it was possible to maintain production of ${\beta}-galactosidase$ activity per cell(specific ${\beta}-galactosidase$ activity) at 14,000, 9600, 45 Miller units in the presence of 1% nitrate. and for a nitrate - independent nar promoter (pNW618) at 12,000, 10,400 and 58 Miller units in the absence of nitrate ion, respectively.

• BMB Reports
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• v.29 no.1
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• pp.92-97
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• 1996

The age-46.5 gene of Escherichia coli is induced by nitrate ion and regulated by Fnr, NarL, and NarP during anaerobic growth. aeg-46.5::lacZ fusion gene shows its maximum expression in narL host after two hours of aerobic to anaerobic switch in M9-Glc-nitrate medium. Fnr and NarP act as positive regulators, and NarL acts as a negative regulator. The control region of the aeg-46.5 was identified and the binding sites of regulator proteins have been predicted (Reznikoff and Choe (1993)). It has two symmetry regions. One is located at -52~-37 bp from the anaerobic mRNA 5'-end, which is the binding site of NarL and NarP. The other is located at +37~+56 bp from the 5'-end of mRNA. In this study, the downstream symmetry region from the mRNA 5'-end was investigated by site-directed mutagenesis. The destruction of the symmetry region increases the expression level of aeg-46.5. We propose that the symmetry region interferes with the expression of aeg-46.5 possibly by forming a stem-and-loop structure.

• Biomolecules & Therapeutics
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• v.23 no.4
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• pp.339-344
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• 2015

Naringenin (NAR) as one of the flavonoids observed in grapefruit has been reported to exhibit an anti-cancer activity. However, more detailed mechanism by which NAR exerts anti-cancer properties still remains unanswered. Thus, in this study, we have shown that NAR down-regulates the level of cyclin D1 in human colorectal cancer cell lines, HCT116 and SW480. NAR inhibited the cell proliferation in HCT116 and SW480 cells and decreased the level of cyclin D1 protein. Inhibition of proteasomal degradation by MG132 blocked NAR-mediated cyclin D1 downregulation and the half-life of cyclin D1 was decreased in the cells treated with NAR. In addition, NAR increased the phosphorylation of cyclin D1 at threonine-286 and a point mutation of threonine-286 to alanine blocked cyclin D1 downregulation by NAR. p38 inactivation attenuated cyclin D1 downregulation by NAR. From these results, we suggest that NAR-mediated cyclin D1 downregulation may result from proteasomal degradation through p38 activation. The current study provides new mechanistic link between NAR, cyclin D1 downregulation and cell growth in human colorectal cancer cells.

• Biomolecules & Therapeutics
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• v.24 no.2
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• pp.140-146
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• 2016

Naringenin (NAR) as one of the flavonoids observed in grapefruit has been reported to exhibit an anti-cancer activity. Activating transcription factor 3 (ATF3) is associated with apoptosis in human colon cancer cells. This study was performed to investigate the molecular mechanism by which NAR stimulates ATF3 expression and apoptosis in human colon cancer cells. NAR reduced the cell viability and induced an apoptosis in human colon cancer cells. ATF3 overexpression increased NAR-mediated cleaved PARP, while ATF3 knockdown attenuated the cleavage of PARP by NAR. NAR increased ATF3 expression in both protein and mRNA level, and increased the luciferase activity of ATF3 promoter in a dose-dependent manner. The responsible region for ATF3 transcriptional activation by NAR is located between -317 and -148 of ATF3 promoter. p38 inhibition blocked NAR-mediated ATF3 expression, its promoter activation and apoptosis. The results suggest that NAR induces apoptosis through p38-dependent ATF3 activation in human colon cancer cells.

• Korean Journal of Animal Reproduction
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• v.13 no.1
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• pp.18-25
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• 1989

This study was carried out to investigate the general semen characteristics of the Korean native goat and the effect of temperature, incubation time, dilution rate, freezing rate and glycerol concentration on motility and NAR (normal apical ridge) acrosome of fresh and frozen Korean native goat spermatozoa. 1. Average semen volume per ejaculate, motility, concentration and pH of fresh Korean native goat spermatozoa were 0.19${\pm}$0.09 ml, 94.5${\pm}$0.47%, 26.17${\times}$108${\pm}$1.68/ml and 6.63${\pm}$0.18, respectively. 2. Motility and NAR acrosome of fresh spermatozoa during incubation were higher at 22$^{\circ}C$ than at 5$^{\circ}C$ or 37$^{\circ}C$(P<.01). 3. Motility and NAR acrosome of spermatozoa diluted 1:4 during incubation were higher at 22$^{\circ}C$ than at 5$^{\circ}C$ or 37$^{\circ}C$(P<.01). 4. Motility and NAR acrosome of spermatozoa during incubation were higher for samples diluted 1:1, 1:2, or 1:4 than for samples diluted 1:6(P<.01). 5. Motility and NAR acrosome of post-thaw spermatozoa were higher at freezing rate of 12$^{\circ}C$/min than at freezing rate of 1$^{\circ}C$/min or 24$^{\circ}C$/min when glycerol concentration was 9%(P<.01).

• KSBB Journal
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• v.11 no.4
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• pp.431-437
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• 1996

The nar promoter of Escherichta coli, which is maximally induced under anaerobic conditions in the presence of nitrate, was characterized to see whether the nar promoter cloned onto pBR322 can be used as an expression promoter. The modified nar promoter, in which several bases in the -10 region was mutated to the consensus sequence by site-directed mutagenesis, was characterized in E. coli, on which chromosome the fnr gene affecting expression of the nar promoter according to dissolved oxygen level was mutated. The E. coli lacZ gene was used as a reporter gene. The following effects were investigated to find optimal conditions to induce the modified nar promoter: induction methods, optimal nitrate concentrations, the amount of ${\beta}$-galactosidase expressed at the different growth conditions, and induction characteristics. The following results were obtained from the experiments : expression of ${\beta}$-galactosidase from the modified nar promoter was not affected much by nitrate concentrations. The maximal specific ${\beta}$-galactosidase activity was obtained when E. coli was grown under aerobic conditions, and then the modified nar promoter was induced at OD600=2.2 under microaerobic conditions (DO=1∼2%), under which conditions the maximal specific ${\beta}$-galactosidase activity was 13,000 Miller units. However, the specific ${\beta}$-galactosidase activity was approximately 6,000 Miller units even before the modified nar promoter was induced. Therefore, the modified nar promoter seemed to be useful when the cloned gene wants to be expressed in E. coli constitutively.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.758-761
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• 2001

In this study, the nar promoter of E. coli was characterized to see whether the nar promoter cloned onto pBBR122 can be used as an expression promoter of gram negative microbes. For this purpose, a plasmid with lacZ gene expressing ${\beta}-galactosidase$ instead of the structural genes of nar operon in a gram negative host strain(Agrobacterium.tumefaciens) was used to simplify an assay of induction of the nar promoter. The following effects were investigated to find optimal conditions: methods of inducing the nar promoter, optimal nitrate concentration, maximally inducing the nar promoter, the amount of expressed ${\beta}-galactosidase$ and induction ratio(specific ${\beta}-galactosidase$ activity after maximal induction/specific ${\beta}-galactosidase$ activity before induction). The following results were obtained from the experiments: the growth of Agrobacterium with E.coli nar promoter was not much affected by nitrate concentration in the shake-flask; induction of nar promoter was optimal when Agrobacterium was grown in the presence of 1% nitrate ion at the beginning of culture and when overnight culture was completely grown in the shake-flask before being transferred to other shake-flask; the amount of ${\beta}-galactosidase$ per cell and per medium volume was maximal when Agrobacterium was grown under aerobic condition to $OD_{600}$ of 1.7; then the nar promoter was induced under microaerobic and anaerobic condition made by lowering dissolved oxygen level(DO). After 2-3h of induction in the YEP medium selected as a main culture medium, the specific ${\beta}-galactosidase$ activity became about 17,000 Miller units in the fermentor cluture.