• Title/Summary/Keyword: Bt. Protein

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Molecular biological analysis of Bt-transgenic (Bt-9) rice and its effect on Daphnia magna feeding

  • Oh, Sung-Dug;Yun, Doh-Won;Chang, Ancheol;Lee, Yu-jin;Lim, Myung-Ho
    • Korean Journal of Agricultural Science
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    • v.46 no.1
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    • pp.113-124
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    • 2019
  • Insect-resistant transgenic (Bt-9) rice was generated by inserting mCry1Ac1, a modified gene from the soil bacterium Bacillus thuringiensis, into the genome of a conventional variety of rice (Ilmi). With regard to potential problems such as safety, an evaluation of non-target organisms is necessary as an essential element of an environmental risk assessment of genetically modified (GM) crops. We studied the effects of the Bt-9 rice on the survival of cantor Daphnia magna, a commonly used model organism in ecotoxicological studies. D. magna fed on the Bt-transgenic rice (Bt-9) and its near non-GM counterparts (Ilmi) grown in the same environment (a 100% ground rice suspension). The Bt-9 rice was confirmed to have the inserted T-DNA and protein expression evident by the PCR and ELISA analyses. The feeding study showed a similar cumulative immobility and abnormal response of the Daphnia magna between the Bt-9 rice and Ilmi. Additionally, the 48 h-EC50 values of the Bt-9 and Ilmi rice were 4,400 mg/L (95% confidence limits: 3861.01 - 5015.01 mg/L) and 5,564 mg/L (95% confidence limits: 4780.03 - 6476.93 mg/L), respectively. The rice NOEC (No observed effect concentration) value for D. magna was suggested to be 1,620 mg/L. We conclude that the tested Bt-9 and Ilmi have a similar cumulative immobility for D. magna, a widely used model organism, and the growth of Bt-9 did not affect non-target insects.

Expression of Rat Hepatic Glutathione-S-Transferases by Benzoazoles (Benzoazole계 화합물이 glutathione-S-transferases의 유도발현에 미치는 영향)

  • 서경원;김연정;김태완;김효정;조민경;김상건
    • Environmental Analysis Health and Toxicology
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    • v.13 no.3_4
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    • pp.55-61
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    • 1998
  • Glutathione-S-transferases (GSTs) detoxify electrophilic xenobiotics and reactive metabolites. Recently benzene-fused heterocycles have been shown to increase the total amount of hepatic GSTs in rats. Primarily this study aimed to determine the induction of GSTs by benzoazoles (BAs) including benzoxazole (BX), 2-methylbenzoxazole (M-BX), 2,5-dimethyl benzoxazole (D-BX), benzothiazole (BT), aminobenzothiazole (A-BT) and 2-mercaptobenzothiazole (M-BT) in rats. Hepatic cytosol and poly(A)$^+$ mRNA were prepared from rats after oral administration of BX, BT, M-BX, D-BX, A-BT and M-BT for 5 consecutive days at doses of 1 mmol/kg. Western immunoblot and northern blot analysis were conducted with rabbit anti-GST Ya, Yb$_1$, Yb$_2$, Yc antibodies and cDNA probes containing = 500 bps in the specific coding regions of Ya, Yb$_1$, Yb$_2$, Yc$_1$, and Yc$_2$, respectively. All BAs increased the amount of enzymes and mRNA levels of GSTs. BT was the most effective inducer of GSTs among the compounds examined in this study. Although A-BT and M-BT, the derivatives of BT, induced GSTs, these chemicals had lesser effect on induction of GSTs than BT. The derivatives of BX also induced less GSTs than the parent compound and the addition of methyl group to the benzene ring of BX reduced the induction of GSTs. BAs had better inductive effects on the class $\alpha$(Ya, Yc) than class $\mu$ GSTs (Yb$_1$, Yb$_2$). BAs enhanced mRNA levels of GSTs in parallel with the protein levels. These results indicate that 1) most of BAs induced various isozymes of GSTs, 2) the induction of GSTs appears to be correlated with the chemical structure of the derivatives, and 3) the expression of GST by BAs is presumably under the transcriptional regulation.

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Betulin Targets Lipin1/2-Meidated P2X7 Receptor as a Therapeutic Approach to Attenuate Lipid Accumulation and Metaflammation

  • Dou, Jia-Yi;Jiang, Yu-Chen;Hu, Zhong-He;Yao, Kun-Chen;Yuan, Ming-Hui;Bao, Xiao-Xue;Zhou, Mei-Jie;Liu, Yue;Li, Zhao-Xu;Lian, Li-Hua;Nan, Ji-Xing;Wu, Yan-Ling
    • Biomolecules & Therapeutics
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    • v.30 no.3
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    • pp.246-256
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    • 2022
  • The present study focused on the potential mechanism of betulin (BT), a pentacyclic triterpenoid isolated from the bark of white birch (Betula pubescens), against chronic alcohol-induced lipid accumulation and metaflammation. AML-12 and RAW 264.7 cells were administered ethanol (EtOH), lipopolysaccharide (LPS) or BT. Male C57BL/6 mice were fed Lieber-DeCarli liquid diets containing 5% EtOH for 4 weeks, followed by single EtOH gavage on the last day and simultaneous treatment with BT (20 or 50 mg/kg) by oral gavage once per day. In vitro, MTT showed that 0-25 mM EtOH and 0-25 µM BT had no toxic effect on AML-12 cells. BT could regulate sterolregulatory-element-binding protein 1 (SREBP1), lipin1/2, P2X7 receptor (P2X7r) and NOD-like receptor family, pyrin domains-containing protein 3 (NLRP3) expressions again EtOH-stimulation. Oil Red O staining also indicated that BT significantly reduced lipid accumulation in EtOH-stimulated AML-12 cells. Lipin1/2 deficiency indicated that BT might mediate lipin1/2 to regulate SREBP1 and P2X7r expression and further alleviate lipid accumulation and inflammation. In vivo, BT significantly alleviated histopathological changes, reduced serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and triglyceride (TG) levels, and regulated lipin1/2, SREBP1, peroxisome proliferator activated receptor α/γ (PPARα/γ) and PGC-1α expression compared with the EtOH group. BT reduced the secretion of inflammatory factors and blocked the P2X7r-NLRP3 signaling pathway. Collectively, BT attenuated lipid accumulation and metaflammation by regulating the lipin1/2-mediated P2X7r signaling pathway.

Construction of a Novel Recombinant Baculovirus Producing Polyhedra with a Bacillus thuringiensis Cry1Ac Crystal Protein

  • Je, Yeon-Ho;Jin, Byung-Rae;Roh, Jong-Yul;Chang, Jin-Hee;Kang, Seok-Kwon
    • The Journal of Korean Society of Virology
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    • v.29 no.3
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    • pp.145-153
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    • 1999
  • We have now constructed a novel recombinant baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV) producing polyhedra with Bacillus thuringiensis (Bt) CryIAc crystal protein. The recombinant polyhedra produced by the recombinant baculovirus, Btrus, in insect cells was characterized. The recombinant baculovirus has two independent transcription units in opposite orientations with two promoters, p10 or polyhedrin gene promoter each initiating transcription of either native polyhedrin or fusion protein with polyhedrin and Bt Cry1Ac crystal protein. Surprisingly, this recombinant baculovirus stably produced recombinant polyhedra which were nearly similar to those of wild-type AcNPV. The immunogold staining experiment showed that the recombinant polyhedra were assembled with polyhedrin and Bt Cry1Ac crystal protein, and contained virus particles. Insecticidal toxicity of recombinant polyhedra of Btrus to the fall webworm, Hyphantria cunea, was strikingly improved in comparison with the wild-type AcNPV.

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Expression and Synergistic Effect of Bacillus thuringiensis CrylAc in Lepidopteran Toxic Strain to Plutella xylostella

  • Kang, Joong-Nam;Roh, Jong-Yul;Shin, Sang-Chul;Ko, Sang-Hyun;Chung, Yeong-Jin;Kim, Yang-Su;Wang, Yong;Choi, Hee-Kyu;Li, Ming-Shun;Choi, Jae-Young;Je, Yeon-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.14 no.1
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    • pp.33-36
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    • 2007
  • To improve insecticidal activity of B. thuringiensis 2385-1 (Bt 2385-1), a recombinant plasmid, pHT1K-1Ac, was introduced into lepidopteran toxic Bt 2385-1 by electroporation. The presence of the recombinant plasmid in Bt 2385-1 after electroporation was confirmed by PCR. Bt 2385-1 transformant was named as Bt pHT1K-1Ac/2385-1 (1K-1Ac/2385-1). The 1K-1Ac/2385-1 transformant produced bipyramidal-shaped parasporal inclusion as like the wild-type strain, Bt 2385-1, and showed an 130 kDa band of Cry1Ac protein. The insecticidal activity of 1K-lAc/2385-1 against S. exigua was similar to that of Bt 2385-1 but the $LC_{50}$ value of transformant against P. xylostella was 1.8 times lower. Through these bioassay results, it was confirmed that toxicity of Bt 2385-1 transformant showed synergistic effect by introducing Cry1Ac. These results suggested that the multiple expressions of Cry proteins in a promising Bt strain may interact synergistically in insect midgut, resulting in increase of toxicity and expansion of host spectrum.

Expression of Bacillus thringiensis HD-1 gene in rhizobacteria Pseudomonas fluorescens KR164 (근권 길항세균 Pseudomonas fluorescens KR164에 Bacillus thuringiensis HD-1 유전자의 삽입과 발현)

  • Kim, Yeong-Yil;Rhee, Young-Hwan;Kang, Heun-Soo
    • Applied Biological Chemistry
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    • v.35 no.4
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    • pp.227-231
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    • 1992
  • The plasmids pSUPBT and pSUPBTR were constructed with a vector pSUP2021 and the BT toxin gene in the plasmid pES 1. The plasmids constructed were introduced into the antagonistic rhizobacteria P. fluorescens KR164 by conjugation and P. fluorescens having pSUPBT and pSUPBTR were named P. fluorescens KR164(pSUPBT)#2, KR164(pSUPBT)#3, KR164(pSUPBTR)#2 and KR164(pSUPBTR)#3, respectively. The BT toxin gene were identified in all transformants by Southern hybridization and the final product of BT toxin gene was identified only in P. fluorescens KR164(pSUPBTR)#3 by SDS-PAGE. This crystal toxin protein were also observed in electron microscopy.

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Evaluation of Dietary Lipid Sources for Juvenile Abalone (Haliotis disucs hannai) (참전복 치패 사료의 지질원 평가)

  • 이상민;박흠기
    • Journal of Aquaculture
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    • v.11 no.3
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    • pp.381-390
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    • 1998
  • Three feeding experiments were carried out to evaluate the effects of fatty acids or lipid sources in diets on the survival, growth and body composition of junenile abalone(Haliotis discus hannai). Diets used in this study contained casein or fish meal as a protein source. Three replicate groups of abalone averaging 160 mg were fed with casein diets containing 12:0, 18:1, 18:2n-6, 18:3n-3, n-3HUFA, squid liver oil (SO), corn oil (CO), beef tallow (BT), SO+CO, and SO+BT, or fed fish meal diets containing SO, CO, BT, SO+CO, SO+BT and not supplemental oil for 20 weeks, respectively. Survival rate, weight gain and soft body weight of abalone were not significantly affected by different fatty acids in the casein diets (P>0.05). Weight gain, soft body weight and shell length of abalone fed the casein diets containing SO, SO+CO or SO+BT were significantly higher (P<0.05) than those of abalone fed the casein diets containing CO or BT. Survival rate of abalone fed the fish meal diets was not influenced by different lipid sources (P>0.05). Weight gain and soft body weight of abalone fed the fish meal diets containing beef tallow (BT or SO+BT diet) were lower than those of abalone fed the diet not added oil or diets containing SO, CO and/or SO+CO(P<0.05). These data indicated that SO or SO+CO was good dietary lipid source for juvenile abalone, and that these oil supplement in diet was not necessary when fish meal was used as a protein source.

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The Effect of $\delta$-endotoxin of Bacillus thuringiensis on the Protein Synthesis of the Several Tissues of Hyphantria cunea Drury (Bacillus thuringiensis 내독소단백질이 미국흰불나방 조직의 단백질 합성에 미치는 영향)

  • 전향미;강석권;서숙재
    • Journal of Sericultural and Entomological Science
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    • v.37 no.1
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    • pp.68-73
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    • 1995
  • The toxic effect of Bacillus thuringiensis var, kurstaki $\delta$-endotoxin was determined in the midgut, fat body and Malpighian lobules from larvae of Hyphantria cunea using the au-toradiography and Western blot methods. Bt $\delta$-endotoxin crystals inhibited protein synthesis and elongation factor-2 activity of all tissues tested.

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