• Journal of Microbiology and Biotechnology
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• v.9 no.5
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• pp.541-547
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• 1999

Constructions of a transfer vector and a recombinant baculovirus using the thymidine kinase gene of the Herpes simplex virus type 1 strain F (HSV -1) were carried out. Newly cloned transfer vector, pHcgXIIIB, was constructed by insertion of the glycoprotein gX gene signal peptide sequence of Pseudorabies virus into the baculovirus vector pHcEV-IV. The gX sequence was inserted just downstream from the promoter for the polyhedrin gene of the Hyphantria cunea nuclear polyhedrosis virus (HcNPV). HSV-1 thymidine kinase(tk) gene (1.131 kb) was used as a candidate gene for transferring into the baculovirus expression system. The tk gene was inserted into a BamHI site downstream from the gX sequence-promoter for the polyhedrin gene in the pHcgXIIIB transfer vector and was transferred into the infectious lacZ-HcNPV expression vector. Recombinant virus was isolated and was named gX-TK-HcNPV. The recombinant virus produced a 45 kDa gX-TK fusion protein in Spodoptera frugiperda cells, which was confirmed by Western blot analysis. Microscopic examination of gX-TK-HcNPV-infected cells revealed normal multiplication. Fluorescent antibody staining indicated that the gX-TK fusion protein was present in the cytoplasm. These results indicated that the transfer vector successfully transferred the gX-tk gene into the baculovirus expression system.

• Biomedical Science Letters
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• v.21 no.2
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• pp.60-68
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• 2015

NecroX-7 is a novel small compound of the NecroX series based on the indole moiety, which has potent cytoprotective and antioxidant properties. We previously detected potential immune regulatory effects of NecroX-7 in immune related diseases like Graft-versus-Host Disease. However, the function and the underlying mechanisms of immunological effects of NecroX-7 in the immune system have not been well established. In this study, we investigated the immune response characterization of differentially expressed genes of NecroX-7 administration in $CD4^+$ T cells by microarray analysis. $CD4^+$ T cells stimulated with NecroX-7 ($40{\mu}M$) or vehicle for 72 hours resulted in the identification of 337 differentially expressed genes (1.5 fold, P<0.05) by expression profiling analysis. Twenty eight of the explored NecroX-7-regulated genes were related to immune system processes. These genes were validated by quantitative real-time PCR. The most significant genes were glutathione reductase, eukaryotic translation elongation factor 1, lymphotoxin-alpha, heat shock protein 9 and chloride intracellular channel protein 4. These findings demonstrate the strongly immune response of NecroX-7 in $CD4^+$ T cells, suggesting that cytoprotection and immune regulation may underlie the critical aspects of NecroX-7 exposure.

• Journal of Nutrition and Health
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• v.12 no.2
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• pp.87-93
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• 1979

인구적 증가와 사료 가격의 상승으로 인한 육류 제품의 가격이 상승하고 있음은 우리나라를 비롯하여 세계적인 현황이다. 이 문제에 대처하는 한 방법으로 새로운 단백질 급원으로써 Single Cell Protein이 개발되었고 이에 대한 여러가지 연구가 되어지고 있다. Single Cell Protein이 인간에게 식용이 될 동물이나 나아가서는 인간에게 직접 식품으로 사용되기 전에 인체나 동물 체내에 끼칠 영향과 그 안전성을 확인하려는 하나의 노력으로 이 연구가 시도되었다. 동물 체내 대사에 미치는 SCP의 악영향의 주원인은 함유된 독성물질로 보고 그 독성물질이 SCP 생산 및 처리 과정 중 어느 시기에 생겨나는가를 규명해 보고자 하였다. Fermentation을 끝낸 SCP의 Supernatant내에 함유된 Inorganic phosphate(I.P.)의 함량을 측정하여 SCP세포의 Viability를 알아보았다. Fermentation을 끝낸 후 10일까지는 I.P.가 차츰줄다가 40일이 지났을 때에는 I.P.의 증가를 보였으나 직후보다는 낮음을 나타냈다. 또한 냉동 건조시킨 뒤에도 SCP Cell이 살아있음을 보였다. 또한 위와 관련지어서 SCP를 Protein급원으로 써서 사육한 흰쥐에 나타난 Protein Efficiency Ratio(PER)를 통하며 SCP의 질을 평가하였다. 실험결과로 나타난 것을 보면 표준 식이에 사용된 Casein의 PER$(\overline{X}=2.58)$이 SCP의 PER$(Dict 1;\overline{X}=0.888$, $Dict 2;\overline{X}=0.893$, $Dict 3;\overline{X}=0.860)$ 보다 유의적으로 높았다. 그리고 총실험기간의 평균치를 볼 때, Fermentation을 끝내고 3일 후와 6일 후에 냉동 건조시킨 SCP의 PER은 시판되는 SCP의 것과 별다른 차이를 보이지 않았다. 그리나 후반기에 있어서는 Fermentation을 끝내고 3일과 6일 후에 냉동 건조시킨 SCP의 PER이 시판되는 SCP의 것보다 약간 저조함을 보였다. 그리고 Casein의 PER은 총 사육기간은 통하여 별 변동이 없음에 반하여 후반기에 SCP의 PER이 급격한 저하를 보임은 SCP 사용상의 문제점을 나타낸 것으로 해석 된다.

• The Korean Journal of Physiology
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• v.3 no.2
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• pp.17-23
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• 1969

Oxygen consumption rate $(QO_2)$ and protein content of liver and kidney of the Ehrlich ascites tumor-bearing mouse were measured from 6th till 14th day after the inoculation of $4{\times}10^6$ Ehrlich ascites tumor cells. The results thus obtained were compared with those of the groups in which; 1) Whole body x-irradiation with 400 r was done to mouse prior to the inoculation of $4{\times}10^6$ Ehrlich ascites tumor cells, 2) Same number of the irradiated tumor cells were inoculated after subjecting the tumor cells to x-irradiation with 400 r or 900 r in vitro, and 3) the normal, and the following results were obtained; 1. $QO_2$ of the liver and kidney of the tumor-bearing mouse were all lower than the normal and a gradual decrease of $QO_2$ in both liver and kidney was noted as the ascites tumor was progressively developing. 2. In the groups where whole body x-irradiation with 400 r was done, or x-irradiation of ascites tumor cells in vitro with either 400 r or 900 r, $QO_2$ of the liver and kidney were lower than the normal, and the pattern of the decrease was similar in the case of the tumor-bearing mouse. 3. Protein contents in all the groups showed lower values than the normal, and the decrease was gradual as the ascites tumor was developing. 4. $QO_2$ and protein levels in the liver were generally lower than those in the kidney. 5. A certain cancerous metabolism was, therefore, noted in the remote organs of Ehrlich ascites tumor-bearing animal.

• Applied Biological Chemistry
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• v.8
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• pp.81-86
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• 1967

Changes in the protein contents of various organs of the germinating soy bean as well as the effect of X-irradiation on the RNA content of cotyledon of it were investigated. 1) The protein content of germinating organs other than cotyledon increase while that of cotyledon decreases as germination proceeds, which is indicative of the fact that the former organs are anabolic and the latter catabolic in nature with regard to their protein metabolism during germination. 2) The cotyledon RNA content of soybean decreases until the 6 th day after germination initiated, and increases thereafter . When the seeds are exposed to $300{\gamma}$ X-irradiation, these tendencies seem to be accelerated, but when exposed to $600{\gamma}$ and $900{\gamma}$ X-irradiation the decrement of RNA in the early stage is prominent and the synthetic recovery in the later period is inhibited markedly due to radiation damage rather than radiation activation.

• International Journal of Industrial Entomology and Biomaterials
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• v.34 no.1
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• pp.1-5
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• 2017

Bone morphogenetic protein 2 (BMP2) plays an important role in the development of bone and cartilage. It is involved in the hedgehog pathway, TGF beta signaling pathway, and in cytokine-cytokine receptor interaction. It is involved also in cardiac cell differentiation and epithelial to mesenchymal transition. In this study, We expressed human BMP2 (hBMP2) recombinant protein using Baculovirus Expression Vector System (BEVS) in Sf9 insect cells. The hBMP2 cDNA was cloned into baculovirus transfer vector, pBacgus-4x-1 and recombinant baculovirus was screened out through X-gal and GUS-fusions assay. Western blot analysis shown that molecular weight of hBMP2 recombinant protein was about 44.71 kDa.

• Journal of the Korean GEO-environmental Society
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• v.13 no.11
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• pp.37-42
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• 2012

This study investigated the use of Geobacter lovleyi with TBOS(Tetrabutoxysilane) for TCE(Trichloroethylene) dechlorination. The TCE dechlorination by Geobacter lovleiy was mathematically described as the independent variables such as initial concentration of TCE, protein mass of Geobacter lovleyi and initial concentration of TOBS, and these were modeled by the use of response surface methodology(RSM). These experiments were carried out as a Box-Behnken Design(BBD) consisting of 15 experiments. The application of RSM yielded the following equation, which is empirical relationship for the dechlorination efficiency($Y_1$, %) of TCE and first order kinetic constant of TCE($Y_2,\;d^{-1}$) by independent variables in coded unit : $Y_1=-11.50X_1$(initial concentration of TCE) + $4.25X_2$(protein mass as Geobacter lovleyi injected mass) - $4.75X_3$(initial concentration of TBOS) - ${6.58X_1}^2$ - ${8.58X_2}^2$ + 93.67, $Y_2=-10.92X_1+5.06X_2-4.89X_3-{4.93X_3}^2-2.19X_1X_2+2.54X_1X_3-2.19X_2X_3+16.71$. In this case, the value of the adjusted determination coefficient(adjusted $R^2$= 0.975 and 0.934) were closed to 1, showing a high significance of the model. Statistical results showed the order of TCE dechlorination at experimental factors to be initial TCE concentration > initial TBOS concentration > protein mass, but the interaction effects were non-significant.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.21 no.3
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• pp.82-93
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• 2008

Objective: Previously, the methanol extracts of the semen of Xanthium strumsrium could involved anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated Raw 264,7 cells, We evaluated the anti-allergic effects of X. strumarium on rat basophilic leukemia (RBL-2H3) cells, Methodes : To investigate the effect of X. strumarium on the phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187-induced RBL-2H3 cells. The effects of X. strumarium on the degranulation and the pro-inflammatory cytokines secretion and expression from RBL-2H3 cells were evaluated with $\beta$-hexosaminidase assay, ELISA, and RT-PCR analysis, In addition, we examined the effects of X. strumarium on nuclear factor (NF)-${\kappa}B$ activation and $I{\kappa}B-\alpha$ degradation using Western blot analysis. Results : X. strumarium inhibited degranulation and secretions and expressions of pro-inflammatory cytokines, such as tumor necrosis factor-alpha ($TNF-\alpha$), interleukin (IL)-4 and cyclooxygenase (COX)-2, on stimulated RBL-2H3 cells, however, X. strumarium not affect cell viability. In stimulated RBL-2H3 cells, the protein expression level of nuclear factor-kappa B (NF-${\kappa}B$) was decreased in the nucleus by X. strumarium. In addition, X. strumarium suppressed the degradation of inhibitory protein $I{\kappa}B-{\alpha}$ protein in RBL-2H3 cells. Conclusion : These results suggest that X. strumarium inhibits the degranulation and secretion of pro-inflammatory cytokines through blockade of NF-${\kappa}B$ activation and I $I{\kappa}B-{\alpha}$ degradation.

• Journal of the East Asian Society of Dietary Life
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• v.15 no.6
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• pp.759-765
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• 2005

This study was carried out to investigate the feeding effects of fermented wild grape by-product on pork meat qualities. The samples consisted of the pork not fed fermented wild grape byproduct(FWG-X) and the pork fed fermented wild grape byproduct(FWG-O). The moisture, crude protein, crude fat and crude ash were not significantly different between samples. The cholesterol and TBARS of FWG-O were lower than those of the FWG-X, and the salt soluble protein extractability of FWG-O was higher than that of the FWG-X(p<0.05). The calorie, cooking loss, water holding capacity, pH and volatile basic nitrogen were not significantly different between FWG-X and FWG-O. The meat colors of the a and b value of FWG-O were higher than those of the FWG-X, and in case of the fat color, the a value of FWG-O was higher than that of the FWG-X. The hardness, springiness, cohesiveness, gumminess, chewiness and shear force were not significantly different between FWG-X and FWG-O. The total amino acid contents of FWG-X and FWG-O were 74.35 and 69.59g/100g protein, respectively, The raw meat color of FWG-O was higher than that of the FWG(p<0.01), and the cooked meat color(p<0.05), taste(p<0.001), flavor(p<0.001), juiciness(p<0.01) and palatability(p<0.01) were superior to those of the FWG-X. This study showed that fermented wild grape by-product decreased the cholesterol content and lipid oxidation with enhancing the sensory score.

• Animal cells and systems
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• v.9 no.3
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• pp.161-171
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• 2005

Integrin-linked kinase 1 (ILK1) regulates several protein kinases, including PKB/Akt kinase and glycogen synthase kinase ${\beta}$. ILK1 is also involved distinctively in the cell morphological and structural functions by interacting with the components of the extracellular matrix or integrin. According to the information of serum- and glucocorticoid-induced kinase 1 (SGK1) substrate specificity (R-X-R-X-X(S/T)-${\phi};{\phi}$ indicates a hydrophobic amino acid), two putative phosphorylation sites, $Thr^{181}\;and\;Ser^{246}$, were found in ILK1. We showed that ILK1 fusion protein and two fluorescein-labeled ILK1 peptides, $FITC-^{174}RTRPRNGTLN^{183}$ and $FITC-^{239}CPRLRIFSHP^{248}$, were phosphorylated by SGK1 in vitro. We also identified that 14-3-3 ${\theta}\;{\varepsilon}\;and\;{\xi}$, among several 143-3 isotypes $({\beta},\;{\gamma},\;{\varepsilon},\;{\eta},\;{\sigma},\;{\theta},\;{\tau}\;and\;{\xi})$ formed protein complex with ILK1 in COS-1 cells. Furthermore, the phosphorylation of $Ser^{246}$ by SGK1 induced the binding with 14-3-3. It was also demonstrated that 14-3-3-bound ILK1 has reduced kinase activity. Thus, these data suggest that SGK1 phosphorylates $Thr^{181}\;and\;Ser^{246}$ of ILK1 and the phosphorylation of its $Ser^{246}$ makes ILK1 bind to 14-3-3, resulting in the inhibition of ILK1 kinase activity.