• BMB Reports
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• v.36 no.3
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• pp.294-298
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• 2003

The pathological effect of the Bacillus thuringiensis Cry $\delta$-endotoxins on susceptible insect larvae had extensive damage on the midgut epithelial cells. In this study, an ex vivo assay was devised for assessing the insecticidal potency of the cloned Cry4B mosquito-larvicidal protein that is expressed in Escherichia coli. Determination of toxicity was carried out by using a cell viability assay on the midguts that were dissected from 5-day old Aedes aegypti mosquito larvae. After incubation with the toxin proteins, the number of viable epithelial cells was determined photometrically by monitoring the quantity of the bioreduced formazan product at 490 nm. The results showed that the 65-kDa trypsin-activated Cry4B toxin exhibited toxic potency ca. 3.5 times higher than the 130-kDa Cry4B protoxin. However, the trypsin-treated products of the non-bioactive Cry4B mutant (R158A) and the lepidopteran-specific Cry1Aa toxin displayed relatively no ex vivo activity on the mosquito-larval midguts. The ex vivo cytotoxicity studies presented here confirms data that was obtained in bioassays.

• BMB Reports
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• v.43 no.6
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• pp.427-431
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• 2010

Cyt2Aa2 is a mosquito-larvicidal protein produced as a 29 kDa crystalline protoxin from Bacillus thuringiensis subsp. darmstadiensis. To become an active toxin, proteolytic processing is required to remove amino acids from its N- and C-termini. This study aims to investigate the functional role of amino acid residues on the N-terminal ${\beta}1$ and C-terminal ${\alpha}F$ of Cyt2Aa2 protoxin. Mutant protoxins were constructed, characterized and compared to the wild type Cyt2Aa2. Protein expression data and SDS-PAGE analysis revealed that substitution at leucine-33 (L33) of ${\beta}1$ has a critical effect on dimer formation and structural stability against proteases. In addition, amino acids N230 and I233-F237 around the C-terminus ${\alpha}F$ demonstrated a crucial role in protecting the protoxin from proteolytic digestion. These results suggested that ${\beta}1$ and ${\alpha}F$ on the Nand C-terminal ends of Cyt2Aa2 protoxin play an important role in the molecular interaction and in maintaining the structural stability of the protoxin.

• The Korean Journal of Pesticide Science
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• v.4 no.1
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• pp.51-58
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• 2000

These studies were carried out to investigate the toxicities of 43 registered insecticides to the sweetpotato whitefly(Bemisia tabaci, B. biotype). Insecticide activities were evaluated by testing systemic action and residual effect in the laboratory, and control efficacy in the greenhouse. All experiments were tested at the recommended concentration(ppm) of each insecticides. Insect growth regulators (IGRs), pyriproxyfen and teflubenzuron showed >95% ovicidal effect. The insecticides that showed >95% larvicidal activity on 3rd nymphal instars were abamectin, acetamiprid, imidacloprid, pyriproxyfen, and acetamiprid+ ethofenprox. Insecticides with >95% adulticidal activity were abamectin, acetamiprid, diazinon, endosulfan, fenitrothion, imidacloprid, methidathion, pirimiphos-methyl, pymetrozine, spinosad, acetamiprid+ ethofenprox, cartap kydrochloride+buprofezin, and fenpropathrin+fenitrothion. Abamectin, acetamiprid, imidacloprid, pyriproxyfen, and acetamiprid+ethofenprox showed both residual effect and systemic activity. In the control efficacy test on B. tabaci, 90% control values were obtained at 1st day after treatment of the insecticides including abamectin, acetamiprid, imidacloprid, pyriproxyfen and acetamiprid+ethofenprox but in pyriproxyfen, 90% control value was reached at 7th day after treatment. These results indicate that abamectin, acetamiprid, imidacloprid, pyriproxyfen and acetamiprid+ethofenprox can be used in control for B. tabaci in field.