• Title/Summary/Keyword: Bt. Protein

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바이오 센서 및 랩온어칩

  • 박유근
    • The Magazine of the IEIE
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    • v.31 no.1
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    • pp.58-72
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    • 2004
  • Smart sensors and biochip technologies have received a great deal of attention in recent years not only because of the enormous potential markets in the healthcare expenditures but more importantly because of its great impact on the quality of human life in the future. Collaborative research among BT (Bio Technologies), IT (Information Technologies) and NT (Nano Technologies) will bring us a new paradigm of the healthcare services. Examples include disease prediction based on the genetic tests, personal medicines, point-of-care analysis, rapid and sensitive infectious disease diagnostics, environmental monitoring for chemical or biological warfares, intelligent drug delivery systems etc. In this report, recent accomplishment in the research area on biosensors, DNA chips, Protein Chips and Lab-on-a-chips are reviewed.

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Leucine Rich Repeat Sequence of the ${\delta}$ Endotoxin Family of Bacillus thuringiensis

  • Vudayagiri, Suvarchala;Jamil, Kaiser
    • BMB Reports
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    • v.33 no.1
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    • pp.89-91
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    • 2000
  • In this investigation we report our search for the presence of Leucine Rich Repeats (LRRs) in various Bacillus thuringiensis (Bt) sub species. Leucine rich repeats are short sequence motifs present in some proteins. The consensus sequence corresponding to the LRR was present in Crystal proteins of Bacillus thuringiensis sub species. This LRR sequence has been predicted to be involved in proteinprotein interactions or receptor binding functions, hence the importance of this study.

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Effect of Feeding Transgenic Cottonseed vis-à-vis Non-transgenic Cottonseed on Haematobiochemical Constituents in Lactating Murrah Buffaloes

  • Singh, Maha;Tiwari, D.P.;Kumar, Anil;Kumar, M. Ravi
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.12
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    • pp.1732-1737
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    • 2003
  • An experiment was conducted to investigate the effect of feeding transgenic cottonseed (Bt.) vis-a-vis non-transgenic (non-Bt.) cottonseed on blood biochemical constituents in lactating Murrah buffaloes. Twenty Murrah buffaloes in mid-lactation were divided into 2 groups of 10 each. Animals of group I were fed with 39.5% non-transgenic cottonseed in concentrate mixture while the same percentage of transgenic (Bt.) cottonseed was included in the concentrate mixture fed to the animals of group II. Animals of both groups were fed with concentrate mixture to support their milk production requirements. Each buffalo was also offered 20 kg mixed green fodder (oats and berseem) and wheat straw ad libitum. The experimental feeding trial lasted for 35 days. There was no significant difference in the dry matter intake between the two groups of buffaloes. All the buffaloes gained body weight, however, the differences were non significant. Total erythrocyte count, hemoglobin content and packed cell volume were $9.27{\pm}0.70${\times}10^6/{\mu}l$, $13.01{\pm}0.60gdl$ and $34.87{\pm}1.47%$, respectively in group I with the corresponding figures of $8.88{\pm}0.33$, $12.99{\pm}0.52$ and $31.08{\pm}1.52$ in group II. The values of total erythrocyte count, haemoglobin content and packed cell volume did not differ significantly between the two groups of buffaloes. The concentration of plasma glucose, serum total proteins, albumin, globulin, triglycerides and high density lipoprotein were non significantly higher in buffaloes fed non-transgenic cottonseed than in buffaloes fed transgenic cottonseed. The cholesterol concentration was significantly (p<0.01) higher in buffaloes of group I ($136.84{\pm}8.40mg/dl$) than in buffaloes of group II ($105.20{\pm}1.85mg/dl$). The serum alkaline phosphotase, glutamic-oxaloacetate transaminase and glutamic-pyruate transaminase activities did not differ significantly between two groups of buffaloes. However, serum glutamic-pyruate transaminase activity was considerably high in buffaloes fed nontransgenic cottonseed as compared to buffaloes fed transgenic cottonseed. Bt. proteins in serum samples of animals of group II were not detected after 35 days of feeding trial. It was concluded that transgenic cottonseed and non-transgenic cottonseed have similar nutritional value without any adverse effects on health status of buffaloes as assessed from haematobiochemical constituents.

Allergenicity Assessment of Cry Proteins in Insect-resistant Genetically Modified Maize Bt11, MON810, and MON863

  • Kim, Jae-Hwan;Seo, Young-Ju;Kim, Ji-Young;Han, Young-Shin;Lee, Kwang-Shin;Kim, Sun-Ah;Kim, Han-Na;Ahn, Kang-Mo;Lee, Sang-Il;Kim, Hae-Yeong
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1273-1278
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    • 2009
  • This study aimed to evaluate the potential allergenicity of Cry proteins in insect-resistant genetically modified (GM) maizes (Bt11, MON810, and MON863) using serum screening tests. Serum samples were obtained from Korean children (0-15 years old) with allergic symptoms who had positive maize-specific IgE. The levels of serum specific IgE was measured by the Phadia ImmunoCAP system and considered as positive when they are 0.35 kU/L or higher. Cry proteins (Cry1Ab in Bt11, mCry1Ab in MON810, and Cry3Bb1 in MON863) were expressed in Escherichia coli and purified for serum screening. The reactivity of purified Cry proteins was confirmed by IgE immunoblots in 50 patients (maize-sensitized patients). There was no reaction between Cry proteins and sera from maize-sensitized patients. Our results suggest that these Cry proteins are not likely to cause allergic reactions. Further studies using more sera from patients with true clinical allergies are needed to evaluate the potential allergenicity of novel proteins in GM maize.

The Effect of Conjugated Linoleic Acid(CLA) Supplemented to Different Fat Sources on Fat Depositions and Stearoyl-CoA Desaturase l(SCDl) Gene Expression in Mice (지방 급원을 달리한 식이에 첨가된 CLA가 지방조직과 Stearoyl-Co A Desaturase 1(SCD 1) 발현에 미치는 영향)

  • Lee, Se-Na;Kang, Keum-Jee
    • The Korean Journal of Food And Nutrition
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    • v.20 no.3
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    • pp.245-252
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    • 2007
  • This study investigated the effects of conjugated linoleic acid(CLA) on the fat deposition, triglyceride levels and the expression of stearoyl-CoA desaturase 1(SCD1) in the livers of male ICR mice that were fed with either soybean oil or beef tallow supplemented with CLA. Mice weighing $25{\sim}30$ g were divided into four groups; soybean oil(SBO), and SBO supplemented with 1% CLA(SBOC), beef tallow(BT) and BT supplemented with 1% CLA(BTC). Each group consisted of 10 mice that were fed the experimental diets for 4 weeks. The experimental diets consisted of 64% carbohydrate, 20% protein, and 16% fat in terms of their contributions to total calories. All other nutrients were identical in the diets. Triglyceride measurements were completed using a kit. Fatty acid compositions were analyzed in the liver using gas chromatography. The levels of SCD1 expression were analyzed by RT-PCR in the liver. No significant differences were found for food intake level, body weight and food efficiency among the experimental groups. However, the weights of epididymal fat pads and plasma triglyceride levels were significantly lower in SBOC and BTC(p<0.05) compared to the SBO and BT groups. These effects were similar in the CLA supplemented groups. The expression level of SCD1 gene and ${\Delta}9$ desaturase index were not significantly different, regardless of the fat used for CLA supplementation. Based on these results, addition of CLA showed decreasing effects on the fat depots weight and the concentration of triglyceride regardless of the fat sources. The SCD1 gene expression and ${\Delta}9$ desaturase index were not influenced by the types of fats with respect to the CLA effects.

Vitamin D Improves Intestinal Barrier Function in Cirrhosis Rats by Upregulating Heme Oxygenase-1 Expression

  • Wang, Peng-fei;Yao, Dan-hua;Hu, Yue-yu;Li, Yousheng
    • Biomolecules & Therapeutics
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    • v.27 no.2
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    • pp.222-230
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    • 2019
  • Intestinal barrier dysfunction always accompanies cirrhosis in patients with advanced liver disease and is an important contributor facilitating bacterial translocation (BT), which has been involved in the pathogenesis of cirrhosis and its complications. Several studies have demonstrated the protective effect of Vitamin D on intestinal barrier function. However, severe cholestasis leads to vitamin D depletion. This study was designed to test whether vitamin D therapy improves intestinal dysfunction in cirrhosis. Rats were subcutaneously injected with 50% sterile $CCl_4$ (a mixture of pure $CCl_4$ and olive oil, 0.3 mL/100 g) twice a week for 6 weeks. Next, $1,25(OH)_2D_3$ ($0.5{\mu}g/100g$) and the vehicle were administered simultaneously with $CCl_4$ to compare the extent of intestinal histologic damage, tight junction protein expression, intestinal barrier function, BT, intestinal proliferation, apoptosis, and enterocyte turnover. Intestinal heme oxygenase-1 (HO-1) expression and oxidative stress were also assessed. We found that vitamin D could maintain intestinal epithelial proliferation and turnover, inhibit intestinal epithelial apoptosis, alleviate structural damage, and prevent BT and intestinal barrier dysfunction. These were achieved partly through restoration of HO-1 and inhibition of oxidative stress. Taken together, our results suggest that vitamin D ameliorated intestinal epithelial turnover and improved the integrity and function of intestinal barrier in $CCl_4$-induced liver cirrhotic rats. HO-1 signaling activation was involved in these above beneficial effects.

Construction of Shuttle Promoter-probe and Expression Vectors for Escherichia coli and Bacillus subtilis, and Expression of B. thuringiensis subsp. kurstaki HD-73 Crystal Protein Gene in the Two Species

  • Park, Seung-Hwan;Koo, Bon-Tag;Shin, Byung-Sik;Kim, Jeong-Il
    • Journal of Microbiology and Biotechnology
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    • v.1 no.1
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    • pp.37-44
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    • 1991
  • A shuttle promoter-probe vector, pEB203, was derived from pBR322, pPL703 and pUB110. Using the vector, a useful DNA fragment, 319 bp EcoRI fragment, having strong promoter activity has been cloned from Bacillus subtills chromosomal DNA. Selection was based on chloramphenicol resistance which is dependent upon the introduction of DNA fragments allowing expression of a chloramphenicol acetyl transferase gene. The nucleotide sequence of the 319 bp fragment has been determined and the putative -35 and -10 region, ribosome binding site, and ATG initiation codon were observed. This promoter was named EB promoter and the resultant plasmid which can be used as an expression vector was named pEBP313. The crystal protein gene from B. thuringiensis subsp. kurstaki HD-73 was cloned downstream from the EB promoter without its own promoter. When the resultant plasmid, pBT313, was introduced into Escherichia coli and B. subtilis, efficient synthesis of crystal protein was observed in both cells, and the cp gene expression in B. subtilis begins early in the vegetative phase. The cell extracts from both clones were toxic to Hyphantria cunea larvae.

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A Safety Evaluation of Genetically Modified Feedstuffs for Livestock Production; the Fate of Transgenic DNA and Proteins

  • Beever, D.E.;Glenn, K.;Phipps, R.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.5
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    • pp.764-772
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    • 2003
  • Two genetic constructs used to confer improved agronomic characteristics, namely herbicide tolerance (HT) in maize and soyabean and insect resistance (Bt) in maize, are considered in respect of feeding to farm livestock, animal performance and the nutritional value and safety of animal products. A review of nucleic acid (DNA) and protein digestion in farm livestock concludes that the frequency of intact transgenic DNA and proteins of GM and non-GM crops being absorbed is minimal/non existent, although there is some evidence of the presence of short fragments of rubisco DNA of non-GM soya in animal tissues. It has been established that feed processing (especially heat) prior to feeding causes significant disruption of plant DNA. Studies with ruminant and non-ruminant farm livestock offered GM feeds demonstrated that animal performance and product composition are unaffected and that there is no evidence of transgenic DNA or proteins of current GM in the products of animals consuming such feeds. On this evidence, current HT and Bt constructs represent no threat to the health of animals, or humans consuming the products of such animals. However as new GM constructs become available it will be necessary to subject these to rigorous evaluation.

Functional characterizations of residues Arg-158 and Tyr-170 of the mosquito-larvicidal Bacillus thuringiensis Cry4Ba

  • Leetachewa, Somphob;Moonsom, Saengduen;Chaisri, Urai;Khomkhum, Narumol;Yoonim, Nonglak;Wang, Ping;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.47 no.10
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    • pp.546-551
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    • 2014
  • The insecticidal activity of Bacillus thuringiensis (Bt) Cry toxins involves toxin stabilization, oligomerization, passage across the peritrophic membrane (PM), binding to midgut receptors and pore-formation. The residues Arg-158 and Tyr-170 have been shown to be crucial for the toxicity of Bt Cry4Ba. We characterized the biological function of these residues. In mosquito larvae, the mutants R158A/E/Q (R158) could hardly penetrate the PM due to a significantly reduced ability to alter PM permeability; the mutant Y170A, however, could pass through the PM, but degraded in the space between the PM and the midgut epithelium. Further characterization by oligomerization demonstrated that Arg-158 mutants failed to form correctly sized high-molecular weight oligomers. This is the first report that Arg-158 plays a role in the formation of Cry4Ba oligomers, which are essential for toxin passage across the PM. Tyr-170, meanwhile, is involved in toxin stabilization in the toxic mechanism of Cry4Ba in mosquito larvae.

Purification and risk assessment of Bacillus thuringiensis Vip3Aa protein against Apis mellifera (Bacillus thuringiensis 유래 Vip3Aa 단백질 순수분리 및 꿀벌 (Apis mellifera)에 대한 위해성평가)

  • Jung, Young Jun;Yoo, Su-Hyang;Lee, Jung Ro
    • Korean Journal of Environmental Biology
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    • v.37 no.4
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    • pp.585-591
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    • 2019
  • Most insect-resistant LMOs have been produced by applying Cry and Vip3Aa proteins. Vip3Aa protein is activated during the vegetative stage of Bacillus thuringensis (Bt) and the inhibitory activity of the Vip3Aa protein against pathogenic attacks from lepidopteran insect species is well known. However, a risk assessment of the Vip3Aa protein compared to the Cry protein has not been conducted in South Korea. This study demonstrates a possible risk assessment method for Vip3Aa protein against honeybees (Apis mellifera). For the risk assessment of the protein, we purified the recombinant Vip3Aa protein in Escherichia coli. The survival rate and symptoms of general intoxication of 4 months honeybees were measured after Vip3Aa exposure. These results indicated that there was no significant difference in the survival rate and the symptom between Vip3Aa and the control buffer. In this study, we established standard methods of Vip3Aa protein purification and oral adult toxicity test using A. mellifera as an LMO risk assessment technique for preserving the natural ecosystem of South Korea.