Effects of the P20 protein from Bacillus thuringiensis israelensis on insecticidal crystal protein Cry4Ba.
The accessory protein P20 from Bacillus thuringiensis israelensis has been defined as an important molecular chaperone for forming crystal Cyt1Aa, and enhancing Cry11Aa and Cry4Aa expression. To investigate its putative role in Cry4Ba delta-endotoxin production and toxicity, a p20 gene was cloned and introduced into B. thuringiensis recombinant strain expressing cry4Ba type gene (cry4BLB). The delta-endotoxin synthesis was enhanced by 262%. The generated inclusions were assayed against third instar larvae of Aedes aegypti. The combination of P20 protein with Cry4BLB delta-endotoxin led to a stable mortality rate of 25% with doses ranging from 0.2 mg l(-1) to 0.6 mg l(-1). Cry4BLB crystals produced in the presence of P20 were much less soluble than those produced by the control strain lacking P20 at pH lower than or equal to 10.5. The observed toxicity perturbation correlates with a decrease of Cry4BLB inclusions solubility. The presence of P20 protein has affected Cry4BLB crystallization and altered greatly its solubility properties. Cry4Ba effectiveness against A. aegypti larvae is related to the solubilization step in larval guts environment.
Biopesticides Team [L.P.I.P.], Center of Biotechnology of Sfax, University of Sfax, P.O. Box 1177, 3018 Sfax, Tunisia.
Jihen
J
Elleuch
Biopesticides Team [L.P.I.P.], Center of Biotechnology of Sfax, University of Sfax, P.O. Box 1177, 3018 Sfax, Tunisia. Electronic address: raida.zribi@cbs.rnrt.tn.
Raida Zribi
RZ
Zghal
Biopesticides Team [L.P.I.P.], Center of Biotechnology of Sfax, University of Sfax, P.O. Box 1177, 3018 Sfax, Tunisia.
Ines
I
Ben Fguira
Institut de Recherche pour le Développement (IRD), UMR MIVEGEC (UM1-UM2-CNRS 5290-IRD 224) Maladies Infectieuses et Vecteurs, Ecologie, Génétique, Evolution et Contrôle, Laboratoire de Lutte contre les Insectes Nuisibles (LIN), Montpellier, France.
Marie Noël
MN
Lacroix
Biopesticides Team [L.P.I.P.], Center of Biotechnology of Sfax, University of Sfax, P.O. Box 1177, 3018 Sfax, Tunisia.
Jihed
J
Suissi
Institut de Recherche pour le Développement (IRD), UMR MIVEGEC (UM1-UM2-CNRS 5290-IRD 224) Maladies Infectieuses et Vecteurs, Ecologie, Génétique, Evolution et Contrôle, Laboratoire de Lutte contre les Insectes Nuisibles (LIN), Montpellier, France.
Fabrice
F
Chandre
Biopesticides Team [L.P.I.P.], Center of Biotechnology of Sfax, University of Sfax, P.O. Box 1177, 3018 Sfax, Tunisia.
Slim
S
Tounsi
Department of Biological & Environmental Sciences, College of Arts Sciences, Qatar University, P.O. Box 2713, Doha, Qatar.
Samir
S
Jaoua
D000330:Aedes / Q000187:drug effects* / Q000254:growth & development; D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics* / Q000378:metabolism; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D003460:Crystallization; D004731:Endotoxins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D015964:Gene Expression Regulation, Bacterial; D006460:Hemolysin Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D007306:Insecticides; D007814:Larva / Q000187:drug effects / Q000254:growth & development; D018832:Molecular Chaperones / Q000235:genetics* / Q000378:metabolism; D012995:Solubility; D014169:Transformation, Bacterial; D019076:Transgenes
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007306:Insecticides; D018832:Molecular Chaperones; C030956:insecticidal crystal protein, Bacillus Thuringiensis
20-kDa chaperone-like protein; Cry4BLB larvicidal activity; Toxin solubilization