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Interactions between molecular chaperone P20 and Cyt2Ba7 toxin in Bacillus thuringiensis

Yongxia Shi, Mujin Tang, Yalin Liao, Wei Xu
doi: https://doi.org/10.1101/129833
Yongxia Shi
1Health Quarantine Lab, Guangdong Inspection and Quarantine Technology Center, Guangzhou, People’s Republic of China 510700
2State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou, People’s Republic of China 510275
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Mujin Tang
2State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou, People’s Republic of China 510275
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Yalin Liao
3School of Veterinary and Life Sciences, Murdoch University, Murdoch, West Australia, Australia 6150
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Wei Xu
3School of Veterinary and Life Sciences, Murdoch University, Murdoch, West Australia, Australia 6150
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  • For correspondence: W.Xu@Murdoch.edu.au
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Abstract

P20 or 20-kilodalton protein is a molecular chaperone protein in Bacillus thuringiensis (Bt) which can increase yields and facilitates crystal formation of various insecticidal crystal proteins (ICPs). In previous studies, a B. thuringiensis insecticidal protein gene, cyt2Ba7, was cloned, expressed but its expression level is very low in B. thuringiensis. In this study, various expression vectors were constructed by incorporating p20 in forward or reverse direction in the upstream of cyt2Ba7 and transformed into a B. thuringiensis acrystalliferous strain 4Q7. The result showed that in the presence of P20, the expression of Cyt2Ba7 was significantly increased. Especially when p20 gene was reversely inserted in the upstream of cyt2Ba7 gene, the expression of Cyt2Ba7 was increased ∼3.2 times meanwhile more and bigger crystals were observed under electron microscopy. By using purified Cyt2Ba7, P20 protein and P20-specific antiserum, immunoblotting and ligand blot analysis demonstrated a strong binding affinity between P20 and Cyt2Ba7. These results reveal that P20 can promote the crystal formation and enhance the expression of Cyt2Ba7 as a molecular chaperone, which can be a powerful tool to boost the ICPs production in B. thuringiensis and help develop more effective insect control strategies.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted April 27, 2017.
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Interactions between molecular chaperone P20 and Cyt2Ba7 toxin in Bacillus thuringiensis
Yongxia Shi, Mujin Tang, Yalin Liao, Wei Xu
bioRxiv 129833; doi: https://doi.org/10.1101/129833
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Interactions between molecular chaperone P20 and Cyt2Ba7 toxin in Bacillus thuringiensis
Yongxia Shi, Mujin Tang, Yalin Liao, Wei Xu
bioRxiv 129833; doi: https://doi.org/10.1101/129833

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