• Title/Summary/Keyword: Bt. Protein

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Analysis of Organic Components and Osteoinductivity in Autogenous Tooth Bone Graft Material

  • Kim, Young-Kyun;Lee, Junho;Kim, Kyung-Wook;Um, In-Woong;Murata, Masaru;Ito, Katsutoshi
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.35 no.6
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    • pp.353-359
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    • 2013
  • Purpose: Extensive research is actively ongoing for development of an ideal bone substitute that meets the gold standard. Tooth was selected as a donor site for evaluation of potentials in bone substitutes based on its similar chemical compositions to alveolar bone. Previous studies have evaluated inorganic components of autogenous tooth bone graft material (AutoBT) and osteoconductivity. In continuation from the previous studies, the current study was conducted for analysis of organic components and evaluation of osteoinductivity of AutoBT. Methods: Forty-six extracted teeth were collected from actual patients (Korea Tooth Bank, R&D Institute). Extracted teeth were processed into AutoBT and implanted in dorsal subcutaneous muscular tissues of 15 athymic mice. Biopsy samples were harvested at two, five, and eight weeks. The Bradford assay, sodium dodecyl sulphate polyacrylamide gradient gel, and western blotting were performed for investigation of organic contents of AutoBT. Results: Histology analyses showed signs of new bone formation as early as two weeks. Results of the Bradford assay indicated the existence of noncollagenous proteins (NCP). 0.29% (2.89 mg/g) of proteins were extracted by weight in the root portion of AutoBT; 0.02% (0.029 mg/g) and 1.79% (17.93 mg/g) of proteins were measured by weight in crown and block-form of AutoBT, respectively. However, recombinant human bone morphogenetic protein-2 was not observed in AutoBT. Conclusion: Within the limitation of the current study, AutoBT induced new bone formation by NCP embedded in dentin.

Effect of insect-resistant genetically engineered (Bt-T) rice and conventional cultivars on the brown planthopper (Nilaparvata lugens Stål)

  • Sung-Dug, Oh;Eun Ji, Bae;Kijong, Lee;Soo-Yun, Park;Myung-Ho, Lim;Doh-Won, Yun;Seong-Kon, Lee;Gang-Seob, Lee;Soon Ki, Park;Jae Kwang, Kim;Sang Jae, Suh
    • Korean Journal of Agricultural Science
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    • v.49 no.3
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    • pp.511-520
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    • 2022
  • Insect-resistant transgenic rice (Bt-T) expresses a toxic protein (mcry1Ac1) derived from the soil bacterium Bacillus thuringiensis found in the rice cultivar Dongjin with an insecticidal property against rice leaf roller (Cnaphalocrocis medinalis). In this study, to investigate the impact of Bt-T on non-target organisms, the feed and oviposition preferences and biological parameters of brown planthopper (Nilaparvata lugens Stål) were comparatively analyzed in four rice cultivars: Dongjin (parent variety), Ilmi (reference cultivar), Chinnong (brown planthopper resistant cultivar) and Bt-T. In the Bt-T and Dongjin cultivars, the feed preferences were 32.4 ± 8.3 and 34.1 ± 6.8%, and the oviposition preferences were 32.5 ± 5.1 and 30.0 ± 5.3% respectively, and there was no statistical significance between these rices. Additionally, in the Bt-T and Dongjin cultivars, the total lifespans from egg to adult were 39.5 ± 6.9 and 40.0 ± 5.8 days, and the weights of adult females were 1.78 ± 0.14 and 1.72 ± 0.16 mg, respectively. Therefore, there was no statistical difference in the biological parameters between these two varieties. Overall, the results indicate that the insect-resistant transgenic rice (Bt-T) did not negatively affect the reproduction and life cycle of brown planthopper, a non-target organism.

Evaluation and Assessment of Biosafety for Bacillus thuringiensis (Bt)-transgenic Rice: Responses of Daphnia magna Fed on Bt-transgenic Rice Variety (해충저항성 Bacillus thuringiensis (Bt) 벼의 환경위해성 평가: 해충저항성 Bt벼가 물벼룩(Daphnia magna)에 미치는 영향)

  • Oh, Sung-Dug;Shin, Hye-Chul;Sohn, Soo-In;Lee, Ki-Jong;Kim, Hyo-Jin;Ryu, Tae-Hun;Lee, Jang-Yong;Park, Beom-Seok;Kweon, Soon-Jong;Suh, Seok-Cheol;Park, Jong-Sug
    • Journal of Applied Biological Chemistry
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    • v.54 no.4
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    • pp.296-302
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    • 2011
  • Insect-resistant transgenic rice was developed by inserting the mCry1Ac1 a modified gene from the soil bacterium Bacillus thuringiensis (Bt). For biosafety assessment, we studied the effects on survival of cantor Daphnia magna, a commonly used as a model organism in ecotoxicological studies. D. magna fed on Bt rice and its near non-genetically modified (GM) counterparts (Nakdong) grown in the same environment (100% ground rice suspension). The Bt rice was comfirmed to have the insertion of T-DNA and protein expression by the polymerase chain reaction and ELISA analysis. Feeding study showed similar cumulative immobility and abnormal response of D. magna between Bt rice and non-GM counterparts. 48 h-$EC_{50}$ values of Bt rice and non-GM rice showed 4,429 and 2,889 mg/L respectively. The rice no observed effect concentration (NOEC) values for D. magna was suggested 1,000 mg/L. We conclude that the tested Bt-rice and Nakdong similar cumulative immobility for D. magna the widely used model organism. We found out that there is strong possibility that the growth of Bt rice didn't affect to non-target insects.

Molecular Characterization of a Novel Vegetative Insecticidal Protein from Bacillus thuringiensis Effective Against Sap-Sucking Insect Pest

  • Sattar, Sampurna;Maiti, Mrinal K.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.9
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    • pp.937-946
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    • 2011
  • Several isolates of Bacillus thuringiensis (Bt) were screened for the vegetative insecticidal protein (Vip) effective against sap-sucking insect pests. Screening results were based on $LC_{50}$ values against cotton aphid (Aphis gossypii), one of the dangerous pests of various crop plants including cotton. Among the isolates, the Bt#BREF24 showed promising results, and upon purification the aphidicidal protein was recognized as a binary toxin. One of the components of this binary toxin was identified by peptide sequencing to be a homolog of Vip2A that has been reported previously in other Bacillus spp. Vip2 belongs to the binary toxin group Vip1-Vip2, and is responsible for the enzymatic activity; and Vip1 is the translocation and receptor binding protein. The two genes encoding the corresponding proteins of the binary toxin, designated as vip2Ae and vip1Ae, were cloned from the Bt#BREF24, sequenced, and heterologously expressed in Escherichia coli. Aphid feeding assay with the recombinant proteins confirmed that these proteins are indeed the two components of the binary toxins, and the presence of both partners is essential for the activity. Aphid specificity of the binary toxin was further verified by ligand blotting experiment, which identified an ~50 kDa receptor in the brush border membrane vesicles of the cotton aphids only, but not in the lepidopteran insects. Our finding holds a promise of its use in future as a candidate gene for developing transgenic crop plants tolerant against sap-sucking insect pests.

Potential Strategies for Prolonging the Usefulness of Bacillus thuringiensis in Engineered Rice

  • Bottrell, D.G.;Aguda, R.M.;Gould, F.L.;Theunis, W.;Demayo, C.G.;Magalit, V.F.
    • Korean journal of applied entomology
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    • v.31 no.3
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    • pp.247-255
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    • 1992
  • A laboratory bioassay that incorporates Bacillus thuringiensis (Bt) purified crystal protein toxins into an artificial diet has identified three toxins, CryIA(b), CryIA(c), and CryIIA, to by effective against the yellow stemborer, Scirpophaga incertulas(Walker). Research is aimed at engineering rice that incorporates genes of one of or more of these toxins so as to mimic the insecticidal action of the insect to Bt. The paper discusses potential strategies for slowing the rate of adaptation that include the use of multiple Bt toxins, promoters that express the toxins only in specific plant tissues at specific times, and mixing transgenic and non-transgenic plants.

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Computational Tridimensional Protein Modeling of Cry1Ab19 Toxin from Bacillus thuringiensis BtX-2

  • Kashyap, S.;Singh, B.D.;Amla, D.V.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.788-792
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    • 2012
  • We report the computational structural simulation of the Cry1Ab19 toxin molecule from B. thuringiensis BtX-2 based on the structure of Cry1Aa1 deduced by x-ray diffraction. Validation results showed that 93.5% of modeled residues are folded in a favorable orientation with a total energy Z-score of -8.32, and the constructed model has an RMSD of only $1.13{\AA}$. The major differences in the presented model are longer loop lengths and shortened sheet components. The overall result supports the hierarchical three-domain structural hypothesis of Cry toxins and will help in better understanding the structural variation within the Cry toxin family along with facilitating the design of domain-swapping experiments aimed at improving the toxicity of native toxins.

Study on the effect of Boyanghwanotang on Thrombosis and Elevated Blood Viscosity (혈전증(血栓症) 및 고점도혈증(高粘度血症)에 관(關)한 보양환오탕(補陽還五湯)의 실험적(實驗的) 연구(硏究))

  • Song, Hyo-Jung
    • The Journal of Internal Korean Medicine
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    • v.10 no.1
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    • pp.137-155
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    • 1989
  • In order investigate the effect of Boyanghwanotang (BT) on thrombosis, Sprague-Dawley rats injected endotoxin into the caudal vein were oral administrated solid extract of BT-182. 4mg/200g (Sample A) and 364.8mg/200g (Sample B), and then observed Platelet, fibrinogen, prothrombin time and FDP. Another group injected hydrocortisone acetate into the muscular rump were adopted to observe the effect of BT on elevated blood Viscosity, administrated Sample A and Sample B, thereafter measured body weight, body temperature, whole blood viscosity, plasma viscosity, hematocrit, RBC, WBC, hemoglobin, glucose, albumin, total protein, triglyceride, cholesterol and prothrombin time. 1. Platelet was increased significantly in Sample B compared with the control group. 2. Fibrinogen of the trial groups compared with the control group was increased, and revealed a significance in Sample B. 3. Prothrombin time was decreased significantly in the trial groups compared with the control group. 4. FDP was decreased in the trial groups compared with the control group. 5. Body weight was increased significantly in Sample B after 7 days, body temperature revealed a significance in the trial groups after 5 days. 6. Whole blood viscosity and plasma viscosity of lower shear rates, both were highly decreased in the trial groupn than the control group. 7. The increase of WBC and decrease of hematocrit, RBC and hemoglobin was showed in the trial groups, and Sample B revealed a significance on the increase of WBC and decrease of RBC. 8. The increase of prothrombin time and decrease of glucose, albumin, total protein, triglyceride and cholesterol was showed in the trial group, and Sample B revealed a Significance on the increase of prothrombin time and decrease of albumin, total protein and triglyceride. 9. As compare with Sample A and Sample B, generally Sample B was highly Significant. According to the above results, it was considered that Boyanghwanotang may be the curative effects for thrombosis and elevated blood viscosity.

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Degradation of the Parasporal Crystal Proteins of Bacillus thuringiensis and Induction of Stress Protein Synthesis in Bt $\delta$-endotoxin Ingested Larvae of Fall Webworm, Hyphantria cunea (Bacillus thuringiensis 의 내독소 단백질의 분해와 흰불나방 섭취유충에서 스트레스 단백질 합성의 유발)

  • 전향미;유용만;강석권;서숙재
    • Korean journal of applied entomology
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    • v.33 no.3
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    • pp.178-183
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    • 1994
  • The prolein components of each crystal toxin of Bacillus thuringiensis kurstaki and alzawai were separated by SDS-PAGE. The major clqistal proteins from both shins were composed of paiypeptides having molecular weights of 130 kd and 64 kd. Digestive mixture of both. toxins with t~ypsin and gut juices shared 62 kd polypeptide which may be major actwe toxh. However, aizawal produced much less amount of 62 kd polypeptide than kurstaki did. On ingestion with Bt &-endotoxin, larvae of Hyphantria cunea developed 45 kd stress protein in the several tissues including fat body, which was induced by heat and cold shock.

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Studies on Development of Antagonistic Microorganism by Cell Fusion - Biological control of disease - ) (세포융합에 의한 신 길항미생물 육종에 관한 연구 - 목초 병해의 생물학적 방제 -)

  • 최기춘;이영환;전우복
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.15 no.1
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    • pp.1-12
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    • 1995
  • This study was to investigate an effective biological control of forage diseases and provide a basic data and a model in improving variety of antagonistic bacteria, with growth promoting effect on forage, through cell fusion. The results obtained were summarized as follows; 1. The antagonistic himbacterium against soil-borne phathogenic fungi Fusarium oxysporum and Rhizoctonia solani was isolated from continuous cropping himsphere soil of forage, and its biological and physiological characteristics were investigated. This bacterium was identified as Bacillus subrilis and named BS 101. Another strain for cell fusion was Bacillus thur ingiensis ssp. kurstaki HD-I(BT 37669) with insecticidal crystal. 2. The auxotropic mutants of BS 101 and BT 37669 were derived after mutagenesis using N-methyl-N'nitro- Nitrosoguanidine(NTG) to give amino acid requirement marker. n e s e auxotropic mutants of BS 101 and BT 37669 were named BS 1013(his-) and BT 69(asp-), respectively. 3. The best protoplast requirement was obtained using DM 3 medium, containing 5% casamino acid, 1 M $MgCI_2$ and 2% bovine semm albumin, to give Fusant 3, 7 and 8. BT toxin gene was not identified with fusants by Southern blotting. However, SDS-PAGE analysis of strains showed various protein patterns among fusants. 4. From the dark culture experiment, growth of forage in inoculated soil with antagonistic bacteria was delayed than that of non-inoculated soil with antagonistic bacteria in each continuous cropping soil and in each sterilized soil. On the other hand, growth duration of forage was different between continuous cropping soil and sterilized soil. 5. Seed germination of Alfalfa, Italian ryegrass and Orchardgrass were significantly improved by inoculation of antagonistic bacteria(p< 0.05).

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Molecular characterization of lepidopteran pest-resistant transgenic rice events expressing synthetic Cry1Ac

  • Lee, Kyeong-Ryeol;Shin, Kong Sik;Suh, Seok Cheol;Kim, Ki Young;Jeon, Yong Hee;Park, Beom Seok;Kim, Ju-Kon;Kweon, Soon-Jong;Lee, Yeon-Hee
    • Plant Biotechnology Reports
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    • v.3 no.4
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    • pp.317-324
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    • 2009
  • The insecticidal toxin gene of Bacillus thuringiensis (Bt) is one of the most commonly used in the development of genetically modified (GM) crops. In this research, we analyzed Bt rice showing lepidopteran pest-resistance. The Bt gene is a synthetic Cry1Ac composed of optimal codons for plants, and the Bt protein is targeted to the chloroplast by a transit peptide. Three Cry1Ac rice events (C103-3, C127-1, and C7-1) were analyzed for molecular characterization. C103-3 contains two copies of T-DNA where the left border (LB) region is truncated. Both C7-1 and C127-1 have a single copy of T-DNA, but a part of the vector backbone DNA is inserted into the genome of C127-1; thus, only C7-1 had intact T-DNA. Progenies of C7-1 crossed with the original cultivar, Nakdong, and double-haploid lines from anther culture of lines crossed with the elite cultivar, Dongjin, were analyzed for T-DNA flanking genomic DNA and genotyping. Results showed that an intact T-DNA region without the vector backbone was inserted into the genome and was stably inherited through generations. The C7-1 homozygous event could be used as breeding material to develop GM rice with pest resistance.