• Journal of Life Science
• /
• v.29 no.10
• /
• pp.1159-1163
• /
• 2019

Although there is a growing interest in male menopause, a phenomenon associated with male hormone depletion, current kits using antibodies to quantify male hormones are expensive. In this study, we constructed an in vitro system for verifying the activity or concentration of male steroid hormones using a transcriptional activity test. A reporter plasmid, pGL2-Neo-ARE-AdE1BTATA, which reacts to testosterone, was constructed. In this plasmid, the ARE-AdE1bTATA sequences can be bounded by the testosterone - androgen receptor complex to express luciferase as a reporter. Then, a stable transfection was performed on the human prostate cancer cell line, LNcap-LN3. The constructed LNcap-LN3/pGL2-Neo-ARE-AdE1BTATA testosterone compound detection (TCD) system showed quantitatively proportional luciferase activities to concentrations of $10^{-13}$ to $10^{-8}M$ of standard testosterone. The established in vitro TCD system will contribute to the development of materials for health/functional foods and drugs as it will be possible to search en masse for testosterone-like or testosterone-inhibiting substances derived from natural materials.

• 한국생물공학회:학술대회논문집
• /
• 2002.04a
• /
• pp.534-537
• /
• 2002

An agarase gene from Bacillus cereus ASK202 was expressed in high levels by E. coli BL21(DE3)/pEBA1 using pET28a(+) vector system with the inducible T7 promoter in the presence of isopropyl- ${\beta}$ -thiogalactopyranoside. The open reading frame encodes 761 amino acid residues with a calculated molecular weight of 83,300 daltons and a potential signal peptide about 36 amino acid residues at the N-terminus. E. coli BL21(DE3)/pEBA1 produce 1280 unit/ ${\ell}$ of agarase. The optimum physical condition for the agarase activity was pH 5.6, and $40^{\circ}C$, respectively. The agarase activity was stable up pH $4.0{\sim}9.0$ and $4{\sim}40^{\circ}C$. The km and maximum rate of metabolism for agar were 0.068mg/$m{\ell}$ and 0.094mg/$m{\ell}{\cdot}min$, respectively.

• Journal of The Korean Society of Agricultural Engineers
• /
• v.60 no.3
• /
• pp.15-25
• /
• 2018

Estimating the reference evapotranspiration is an important factor to consider in irrigation system design and agricultural water use. However, there is a limitation in using the FAO Penman-Monteith (FAO P-M) equation, which requires various meteorological data. The purpose of this study is to compare three reference evapotranspiration (ETo) equations in the case of meteorological data missing for 11 study weather stations. Firstly, the FAO P-M equation is used for reference potential evapotranspiration estimation with the actual solar radiation data $R_n$ and the actual vapor pressure $e_a$. Then, in the case of $R_n$, and $e_a$ are missed, the reference evapotranspirations applying FAO P-M, Priestley-Taylor (P-T), Hargreaves (HG) equation were calculated using other meteorological factors. Secondly, MAE, RMSE, $R^2$ were calculated to compare ETo relationship from the ETo equations. From the results, ETo with Hargreaves equation in coastal areas and the Priestley-Taylor equation in the inland areas showed relatively high correlation with FAO P-M when $e_a$ data is missed. In the case of $R_n$ data is missed or two weather data, $e_a$, and $R_n$ data are all missed, $R^2$ value in Priestley-Taylor equation was highest in coastal areas, and $R^2$ values in Hargreaves equation were the high values for 7 inland areas. The results of sensitivity analysis showed that net radiation was the most sensitive for P-T and HG equation, and for FAO P-M, the most sensitive factor was net radiation and relative humidity, air temperature and wind speed were follows. Therefore, in considering of the accessibility to the coast, the types of the missing wether data, and the correlation and the magnitude of error, the reference evapotranspiration equations would be selected in sense of different conditions.

• Microbiology and Biotechnology Letters
• /
• v.51 no.1
• /
• pp.37-42
• /
• 2023

Neutral and non-essential amino acid, glutamine (Gln), plays an essential role in supplying nitrogen to all the amino acids and nucleotides in the mammalian body. Gln is also the most important carbon source that provides intermediates for gluconeogenesis and fatty acid synthesis and supplements the tricarboxylic acid cycle in fast-growing cancer cells. Among the known 14 Gln transporter genes, soluted carrier family 1 member 5 (SLC1A5) has been reported to be closely associated with cancer cell growth. Three variants (v1, v2, and v3) have been derived from SLC1A5. Here, we established a heterologous gene expression system for the active form of human SLC1A5 variant-2 (hSLC1A5v2) in Escherichia coli. v2 is the smallest variant that has not yet been studied. Four expression systems were investigated: pBAD, pCold, pET, and pQE. We also addressed the problem of codon usage bias. Although pCold and pET overexpressed hSLC1A5v2 in E. coli, they were functionally inactive. hSLC1A5v2 using the pBAD system was able to catalyze the successful transport of Gln, even if it was not highly expressed. Initial activity of hSLC1A5v2 for [¹⁴C] Gln uptake in E. coli reached up to 6.73 μmole·min^-1·gDW^-1 when the cell was induced with 80 mM L-arabinose. In this study, we demonstrated a heterologous expression system for the human membrane protein, SLC1A5, in E. coli. Our results can be used for the functional comparison of SLC1A5 variants (v1, v2, and v3) in future studies, to facilitae the developement of SLC1A5 inhibitors as effective anticancer drugs.

• Carbon letters
• /
• v.13 no.3
• /
• pp.173-177
• /
• 2012

A drug delivery system (DDS) was prepared with a temperature and pH-responsive hydrogel. Poly(vinyl alcohol) (PVA)/poly(acrylic acid) (PAAc)/poly(N-isopropylacrylamide) (PNIPAAm)/multi-walled carbon nanotube (MWCNT) nanocomposites were prepared by radical polymerization for the temperature and pH-responsive hydrogels. MWCNTs were employed to improve both the thermal conductivity and mechanical properties of the PVA/PAAc/PNIPAAm/MWCNT nanocomposite hydrogels. Various amounts of MWCNTs (0, 0.5, 1 and 3 wt%) were added to the nanocomposite hydrogels. PVA/PAAc/PNIPAAm/MWCNT nanocomposite hydrogels were characterized with a scanning electron microscope. The mechanical properties were measured with a universal testing machine. Swelling and releasing properties of nanocomposite hydrogels were investigated at various temperatures and pHs. Temperature and pH-responsive release behavior was found to be dependent on the content of MWCNTs in nanocomposite hydrogels.

• Journal of Microbiology
• /
• v.40 no.2
• /
• pp.125-128
• /
• 2002

The aerobic respiratory chain of Vibrio alginolyticus possesses two different kinds of NADH oxidase systems, i.e., an $Na^{+}$-dependent NADH oxidase system and an $Na^{+}$-independent NADH oxidase system. When deamino-NADH, which is the only substrate for the $Na^{+}$-dependent NADH oxidase system, was used as a substrate, the maximum activities of $N^{+}$-dependent NADH: quinone oxidoreductase and $Na^{+}$-dependent NADH oxidase were obtained at about 0.06 M and 0.2 M NaCl, respectively. When NADH, which is a substrate for both $Na^{+}$-dependent and $Na^{+}$-independent NADH oxidase systems was used as a substrate, the NADH oxidase activity had a pH optimum at about 8.0. In cGntrastl when deamino-NADH was used as a substrate, the NADH oxidase activity had a pH optimum at about 9.0. On the other handle inside-out membrane vesicles prepared from the wild-type bacterium generated only a very small $\Delta$pH by the NADH oxidase system, whereas inside-out membrane vesicles prepared from Napl, which is a mutant defective in the $Na^{+}$ pump, generated $\Delta$pH to a considerable extent by the NADH oxidase system. On the basis of the results\ulcorner it was concluded that the respiratory chain-linked components of V. atginotyticus affect each other.

• Proceedings of KOSOMES biannual meeting
• /
• 2018.06a
• /
• pp.194-194
• /
• 2018

• Journal of Microbiology and Biotechnology
• /
• v.20 no.3
• /
• pp.550-557
• /
• 2010

Escherichia coli (E. coli) heat-labile enterotoxin B subunit (LTB) was regarded as one of the most powerful mucosal immunoadjuvants eliciting strong immunoresponse to coadministered antigens. In the research, the high-level secretory expression of functional LTB was achieved in P. pastoris through high-density fermentation in a 5-1 fermentor. Meanwhile, the protein was expressed in E. coli by the way of inclusion body, although the gene was cloned from E. coli. Some positive yeast and E. coli transformants were obtained respectively by a series of screenings and identifications. Fusion proteins LTB-6$\times$His could be secreted into the supernatant of the medium after the recombinant P. pastoris was induced by 0.5% (v/v) methanol at $30^{\circ}C$, whereas E. coli transformants expressed target protein in inclusion body after being induced by 1 mM IPTG at $37^{\circ}C$. The expression level increased dramatically to 250-300 mg/l supernatant of fermentation in the former and 80-100 mg/l in the latter. The LTB-6$\times$His were purified to 95% purity by affinity chromatography and characterized by SDS-PAGE and Western blot. Adjuvant activity of target protein was analyzed by binding ability with GMI gangliosides. The MW of LTB-6$\times$His expressed in P. pastoris was greater than that in E. coli, which was equal to the expected 11 kDa, possibly resulted from glycosylation by P. pastoris that would enhance the immunogenicity of co-administered antigens. These data demonstrated that P. pastoris producing heterologous LTB has significant advantages in higher expression level and in adjuvant activity compared with the homologous E. coli system.

• Proceedings of the KIEE Conference
• /
• 2005.07b
• /
• pp.1123-1125
• /
• 2005

본 논문에서는 3차원 공간에 종이와 같은 박막형 유전체가 있을 때 임의의 점에서의 전계$\vec{E}$를 구하기 위해 유전체의 분극벡터$\vec{P}$를 미지수로 하는 경계적분법(Boundary integral method)을 사용한다. 경계적분법의 사용으로 FEM 3차원 해석에 있어서의 요소분할의 난이성 및 계수 행렬의 대형화로 인한 컴퓨터 수행능력의 한계를 극복할 수 있다. 여기서 분극벡터$\vec{P}$를 구하기 위해 전하에서의 전계${ves{E}}_s$에 의한 유전체내의 분극벡터$\vec{P}$를 수식으로 정리하여 $[\vec{K}][\vec{P}]=[\vec{E}]$ 형태의 $\vec{P}$를 미지수로 하는 system matrix를 구성한다. 위의 system matrix 통해 구해진 분극벡터 $\vec{P}$를 이용하여 유전체 밖의 한 점에서의 전계세기 ${\vec{E}}_m$를 구한 후 우리가 구하고자 하는 전계$\vec{E}$를 계산한다.

• Korean Journal of Veterinary Research
• /
• v.40 no.2
• /
• pp.403-414
• /
• 2000

To improve the semen production, the selenium(Se) and vitamin E(Vit. E) were administrated into Hanwoo young sire for intensification an antioxidant system and the taurine were supplemented into semen extender for improving the semen characteristics. The 16 heads ranging from twenty to thirty two months of age were randomly assigned to control group, Se-admistrated group(Se-group), Vit. E-administrated group(Vit. E-Group) and Se and Vit. E administrated group(Se and Vit. E-group). Se and Vit. E dministrated 3 times every 30 days by intramuscular injection. The administration of Se, Vit. E, and Se and Vit. E didn't affect on semen volume, sperm concentration, and total sperm number among all groups(p>0.05). Adiministration of Se improved sperm motility and viability. Motility of Se-group and control were 26.01% and 19.20%, respectively(p<0.05). Viability of Se-group(47.07%) was higher than control group(35.73%), Vit. E group(36.55%)(p<0.05). The administration of Se and Vit. E didn't affect sperm capacitation and acrosome reaction. The 100mM taurine supplement into semen extender increased the motility of frozen/thawed semen in the Vit. E-group(p<0.05) and had a beneficial effects on decreasing abnormality of frozen/thawed semen in all groups(p<0.05). These results indicate that Se administration improve sperm motility and viability, and the taurine supplement into semen extender decrease abnormality in Hanwoo young sire.