• Title/Summary/Keyword: Bt. Protein

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Protective effect of Buddha's Temple extract against tert-butyl hydroperoxide stimulation-induced oxidative stress in DF-1 cells

  • Eun Hye Park;Sung-Jo Kim
    • Animal Bioscience
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    • v.36 no.7
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    • pp.1120-1129
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    • 2023
  • Objective: This study aimed to determine the protective efficacy of Buddha's Temple (BT) extract against tert-butyl hydroperoxide (t-BHP)-induced oxidative stress in Gallus gallus chicken embryo fibroblast cell line (DF-1) and its effects on the cell lipid metabolism. Methods: In this experimental study, Gallus gallus DF-1 fibroblast cells were pretreated with BT 10-7 for 24 hours, followed by their six-hour exposure to t-BHP (100 μM). Water-soluble tetrazolium salt-8 (WST-8) assays were performed, and the growth curve was computed. The intracellular gene expression changes caused by BT extract were confirmed through quantitative polymerase chain reaction (qPCR). Flow cytometry, oil red O staining experiment, and thin-layer chromatography were performed for the detection of intracellular metabolic mechanism changes. Results: The WST-8 assay results showed that the BT pretreatment of Gallus gallus DF-1 fibroblast cell increased their cell survival rate by 1.08%±0.04%, decreased the reactive oxygen species (ROS) level by 0.93%±0.12% even after exposure to oxidants, and stabilized mitochondrial activity by 1.37%±0.36%. In addition, qPCR results confirmed that the gene expression levels of tumor necrosis factor α (TNFα), TIR domain-containing adapter inducing IFN-beta (TICAM1), and glucose-regulated protein 78 (GRP78) were regulated, which contributed to cell stabilization. Thin-layer chromatography and oil red O analyses showed a clear decrease in the contents of lipid metabolites such as triacylglycerol and free fatty acids. Conclusion: In this study, we confirmed that the examined BT extract exerted selective protective effects on Gallus gallus DF-1 fibroblast cells against cell damage caused by t-BHP, which is a strong oxidative inducer. Furthermore, we established that this extract significantly reduced the intracellular ROS accumulation due to oxidative stress, which contributes to an increase in poultry production and higher incomes.

Stable Transmission and Expression of TPO Transgene up to 10 Generation in the Transgenic Mice (형질전환 생쥐에서 제10세대까지 TPO 유전자의 안정적 전이와 지속적인 발현)

  • 정진우;오건봉;한용만;이경광
    • Korean Journal of Animal Reproduction
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    • v.27 no.1
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    • pp.9-14
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    • 2003
  • The pBT-L transgenic mice carrying human TPO gene in conjunction with bovine $\beta$-casein promoter express human TPO in milk during lactation. In this study, stability of germ line transmission and expression of pBT-L transgene integrated into host chromosome were monitored up to generation F10 of transgenic pBT-L/15 line. When male mouse of generation F8 was crossbred with normal females, approximately half of offsprings (51.3$\pm$18.98%) were identified as transgenic mice. Generation F9 and F 10 mice also showed similar transmission rates (43.8$\pm$18.98% and 71.4$\pm$26.98%, respectively), implying that pBT-L transgene can be transmitted stably up to long term generation in the transgenic mice. Expression levels of human TPO from milk of generation F9 and F10 mice were 1.1$\pm$0.33 mg/ml and 1.1 $\pm$0.45 mg/ml, respectively, which are similar to expression level of generation F2 mice. In conclusion, our results suggest that transgenic animals once established will continuously pass their transgenes to the progeny through the breeding program with the same productivity of human protein in their milk.

Effects of Dietary Lipid Sources on the Growth and Body Composition of the far Eastern Catfish, Silurus asotus (사료 지질원이 메기 Silurus asotus의 성장 및 체조성에 미치는 영향)

  • Kim, Kyoung-Duck;Kim, Jin-Do;Lim, Sang-Gu;Kang, Yong-Jin;Son, Maeng-Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.5
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    • pp.445-450
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    • 2010
  • This study investigated the effects of dietary lipid sources on growth performance and body composition of juvenile far eastern catfish, Silurus asotus. Three replicate groups of fish (average weight 3.6 g) were fed with one of the following experimental diets containing 10% beef tallow (BT), 5% BT plus 5% corn oil (CO), 5% BT plus 5% linseed oil (LO), or 5% BT plus 5% squid liver oil (SO) as the lipid source for 5 weeks. No significant difference was observed in the survival among groups. The weight gain of fish fed the LO (high in 18:3n-3) and SO (high in n-3 highly unsaturated fatty acid) diets was significantly higher than that of the fish fed the CO (high in 18:2n-6) and BT diets (P<0.05). The feed efficiency of fish fed LO and SO diets was significantly higher than that of the fish fed the BT diet (P<0.05), but not significantly different from that of the fish fed the CO diet. The protein efficiency ratio of fish fed the SO diet was significantly higher than that of fish fed the CO and BT diets (P<0.05), but not significantly different from that of fish fed the LO diet. The 18:1n-9 of whole-body polar lipid fraction in fish fed the BT diet increased compared to that of fish fed the other diets. Fish fed the CO and LO diets had significantly higher contents of 18:2n-6 and 20:4n-6, and 18:3n-3, than the fish fed the other diets in polar and non-polar lipid fractions, respectively (P<0.05). Significantly higher contents of 20:5n-3 and 22:6n-3 were observed in the whole-body polar lipid fraction of fish fed the SO diet compared with fish fed the other diets (P<0.05). The study results indicate that linseed oil and squid liver oil containing n-3 fatty acids are good dietary lipid sources for the growth of far eastern catfish.

Molecular Clonging and Hyperexpression of a Bt Gene, cryIAc, in Escherichia coli $DH5{\alpha}$: Production and Usage of Anti-CryIAc Antibody

  • RYOU, CHONGSUK;TAEYOUNG CHUNG;MOOSIK KWON
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1093-1098
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    • 2001
  • The gene coding for a Lepidoptera-specific insecticidal crystalline (or control) protein (ICP), recognized as cryIAc, from Bacillus thuringiensis subsp. kurstaki HD-73, was cloned into the vector pBluscript ll SK-, and then transformed in Escherichia coli $DH5{\alpha}$. The clone was named EBtIAc and the chimeric phagemid, as pEBtIAc. Hyperexpression of CryIAc protoxin was observed in the extract of the culture of E. coli harboring pEBtIAc. Crystalline protoxin was purified by differential solubility. It was dissolved in alkaline pH, and exposed to trypsin to be activated. The molecular weights of the pro- and activated toxins on SDS-PAGE were estimated to be ca. 130 kDa and 60 kDa, respectively. The toxicity was tested by force-feeding larvae of gypsi moth (Lymantria diapar) with trypsinized protoxin. Using the batch of biologically active form of the toxin as an immunogen, anti-CryIAc antiserum was raised in a New Zealand white rabbit. Immunoglobulin G was fractionated from the seam by Protein-A sepharose affinity chromatography. Immunoreactivity of the antibody was examined by dot and Westerns blottings. It has been found that the anti- CryIAc antibody recognized the purified toxin at a level below a nanogram in terms of quantity. Using the antibody some of Bt-corns were able to be differentiated from tons of corn kernels which were imported from America as forage crops.

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Proteomic Analysis of Serum of Women with Elevated Ca-125 to Differentiate Malignant from Benign Ovarian Tumors

  • Li, Li;Xu, Yi;Yu, Chun-Xia
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3265-3270
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    • 2012
  • Clinically, elevated cancer antigen 125 (CA-125) in blood predicts tumor burden in a woman's body, especially in the ovary, but cannot differentiate between malignant or benign. We here used intensive modern proteomic approaches to identify predictive proteins in the serum of women with elevated CA-125 to differentiate malignant from benign ovarian tumors. We identified differentially expressed proteins in serum samples of ovarian cancer (OC) patients, benign ovarian tumor (BT) patients, and healthy control women using mass spectrometry-based quantitative proteomics. Both the OC and BT patients had elevated CA-125. Quantitation was achieved using isobaric tags for relative and absolute quantitation. We obtained 124 quantified differential serum proteins in OC compared with BT. Two proteins, apolipoprotein A-4 (APOA4) and natural resistance-associated macrophage 1, were verified using Western blotting. Proteome profiling applied to OC cases identified several differential serum proteins in the serum of women with elevated CA-125. A novel protein, APOA4, has the potential to be a marker for malignant tumor differentiation in the serum of women with elevated CA-125.

Scanning Electron Microscopy of the Tisues of Helicoverpa assulta Larvae intoxicated with Bacillus thuringiensis Protein Crystals.

  • Cheon Hyang Mi;You
    • Journal of Sericultural and Entomological Science
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    • v.36 no.2
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    • pp.162-167
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    • 1994
  • Surface changes of tissues caused by B. thuringiensis var. kurstaki-$\delta$-endotoxin intoxication of Helicoverpa assulta were observed by scanning electron microscopy. Bt-endotoxin crystals induced the erosion and disruption on the surface of all tissues tested. The results revealed that the toxin binds to all exposed plasma membranes without apparent specificity for particular membrane domains.

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Construction of Transfer Vector for Production of Baculovirus Occlusion Bodies that Contain Novel Recombinant Crystal Protein

  • Shim, Hee-Jin;Choi, Jae-Young;Roh, Jong-Yul;Li, Ming Shun;Je, Yeon Ho
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.118-119
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    • 2003
  • Baculovirus occlusion bodies have been recently engineered to incorporate foreign protein such as the Bacillus thuringiensis (Bt) CrylAc protein for improvement of insecticidal activity. In this study, polyhedrin, cylAc, egfp and crylCa genes were fused to produce occlusion bodies that contain novel recombinant crystal protein by homologous recombination between cylAc and crylCa genes in insect cells. (omitted)

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Genuine traditional Korean medicine, BaekJeol-Tang for the treatment of rheumatoid arthritis

  • Han, Na-Ra;Sim, Woo-Moon;Sul, Moo-Chang;Kim, Min-Cheol;Lee, Chang-Hee;Kim, Dong-Won;Lee, Se-Hun;Lee, Ho-Cheol;Ryu, Jong-Min;Nam, Bong-Soo;Kim, Jong-Ok;Moon, Seong-Oh;Jang, Hyeon-Lok;Kim, Young-Seok;Lee, Ihn;Yang, Jin-Young;Hwang, Kyu-Sun;Chun, Chang-Sun;Jeong, Hyeon-Seok
    • CELLMED
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    • v.3 no.2
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    • pp.18.1-18.7
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    • 2013
  • Inflammation in rheumatoid arthritis is characterized by immune cell infiltration and cytokine secretion. In particular, mast cells and their cytokines play an important role in the pathogenesis of rheumatoid arthritis. Korean medicine, BaekJeol-Tang (BT) was designed by traditional Korean medicine theory. We already reported therapeutic effect of BT in rheumatoid arthritis. Here, we report the specific underlying mechanism of BT in activated human mast cells, HMC-1 cells. In addition, we report for the first time that BT significantly inhibited the production and mRNA expression of proinflammatory cytokines including thymic stromal lymphopoietin, interleukin (IL)-$1{\beta}$, IL-6, IL-8, and tumor necrosis factor-${\alpha}$ in activated HMC-1 cells. BT also decreased the activation of mitogen-activated protein kinases, nuclear factor-${\kappa}B$, and caspapase-1. Taken together, these results indicate that BT has potential as a regulator of inflammatory reactions for the treatment of arthritis such as osteoarthritis and rheumatoid arthritis.

An In Vitro Method to Estimate Apparent Ileal Crude Protein Digestibility in Feed Ingredients Fed to Broiler Chickens (육계에서의 외관상 회장 조단백질 소화율 추정을 위한 In Vitro 실험방법)

  • Su Hyun, An;Changsu, Kong
    • Korean Journal of Poultry Science
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    • v.49 no.4
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    • pp.273-279
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    • 2022
  • The objective of this study was to evaluate an in vitro procedure to estimate the crude protein (CP) digestibility of feed ingredients and mixed diets in broiler chickens. The apparent ileal digestibility (AID) of the CP was measured using 23-d-old male broilers. Three experimental diets, containing three feed ingredients, namely soybean meal (SBM), canola meal (CM), and corn distiller's dried grains with solubles (DDGS), were used as the sole source of CP. A 2-step in vitro procedure was used to estimate in vivo CP digestibility; all the experiments were performed in triplicate. In step 1, the feed ingredient and mixed diet samples were incubated for 4 h at 40 °C with a pH 2.0 pepsin solution, and in step 2, the flasks were incubated for 12 h at 40 °C with a pH 6.8 pancreatin solution. Following incubation, all the samples were filtered; the undigested residues were collected and pooled together to analyze the undigested CP concentration. The in vitro CP digestibility of mixed diets and feed ingredients were 93.2% and 93.0% for SBM, 86.8% and 86.7% for CM, and 83.8% and 79.1% for DDGS, respectively. The coefficients of determination (R2) between the CP digestibility values for the feed ingredients and the in vitro CP digestibility values for the feed ingredients or respective mixed diets were 0.87 or 0.67. The results of the study demonstrated that the in vitro CP digestibility values obtained from the respective mixed diets were better estimates than the values obtained from the individual feed ingredients to predict the AID of CP in feed ingredients fed to broiler chickens.

Expression of the Bacillus thuringiensis Crystal Protein Gene in Pseudomonas Isolated from Rhizosphere Soil of Korean Crops (국내 농작물의 근부토양에서 분리한 Pseudomonas 내에서의 Bacillus thuringiensis 독소단백질 유전자의 발현)

  • Tag, Koo-Bon;Shin, Byung-Sik;Park, Seung-Hwan;Park, Ho-Yong;Kim, Jeong-Il
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.295-300
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    • 1989
  • Screening of Pseudomonas strains that can be used as hosts for expression of crystal protein gene of Bacillus thuringiensis subsp. kurstaki HD-73 was carried out. From rhizosphere soil of 7 kinds or crops as fluorescent Pseudomonas strains were isolated. A hybrie plasmid, pKTC1, composed of the broad host range vector pKT230 and the crystal protein gene was constructed and used for transformation of the 35 Pseudomonas strains. As the result, the crystal protein gene could be introduced into 4 isolates. Several methods including bioassay and immunochemical detection indicated that the crystall protein gene was expressed in the Pseudomonus isolates.

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