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. 2019 Jun 25;12(1):319.
doi: 10.1186/s13071-019-3560-2.

Cloning, expression and activity of ATP-binding protein in Bacillus thuringiensis toxicity modulation against Aedes aegypti

Guo-Hui Zhao  1 , Jian-Nan Liu  1 , Xiao-Hua Hu  1 , Khadija Batool  1 , Liang Jin  1 , Chen-Xu Wu  1 , Juan Wu  1 , Hong Chen  1 , Xiao-Yan Jiang  1 , Zhao-Hui Yang  1 , Xian-Hui Huang  1 , En-Jiong Huang  2 , Xiao-Qiang Yu  3   4 , Xiong Guan  1 , Ling-Ling Zhang  5
Affiliations

Cloning, expression and activity of ATP-binding protein in Bacillus thuringiensis toxicity modulation against Aedes aegypti

Guo-Hui Zhao et al. Parasit Vectors. .

Abstract

Background: Bacillus thuringiensis israelensis (Bti) is a widely used mosquitocidal microbial pesticide due to its high toxicity. ATP-binding proteins (ABP) are prevalently detected in insects and are related to reaction against Bti toxins. However, the function of ABP in mosquito biocontrol is little known, especially in Aedes aegypti. Therefore, this study aimed to clarify the function of ABP in Ae. aegypti against Bti toxin.

Results: Aedes aegypti ABP (GenBank: XM_001661856.2) was cloned, expressed and purified in this study. Far-western blotting and ELISA were also carried out to confirm the interaction between ABP and Cry11Aa. A bioassay of Cry11Aa was performed both in the presence and absence of ABP, which showed that the mortality of Ae. aegypti is increased with an increase in ABP.

Conclusions: Our results suggest that ABP in Ae. aegypti can modulate the toxicity of Cry11Aa toxin to mosquitoes by binding to Bti toxin. This could not only enrich the mechanism of Bt toxin, but also provide more data for the biocontrol of this transmission vector.

Keywords: ATP-binding protein; Aedes aegypti; Biocontrol; Bti; Interaction.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
PCR product of ATP-binding protein. Lane M: 100 bp plus DNA ladder; Lane 1: PCR product of ATP-binding protein
Fig. 2
Fig. 2
Restriction enzyme digestion of pMD-atp recombinant plasmid. Lane M: 100 bp plus DNA ladder; Lane 1: NdeI digested pMD-atp recombinant plasmid; Lane 2: SalI digested pMD-atp recombinant plasmid; Lane 3: NdeI and SalI simultaneously-digested pMD-atp recombinant plasmid
Fig. 3
Fig. 3
SDS-PAGE analysis purified Cry11Aa and ATP-binding protein. a SDS-PAGE analysis of the expressed protein (Lane M: protein molecular weight markers; Lane 1: purified Cry11Aa). b SDS-PAGE analysis of the expressed protein (Lane M: protein molecular weight markers; Lane 1: purified ATP-binding protein)
Fig. 4
Fig. 4
Western Blot analysis of Cry 11Aa and ATP-binding protein. a Western blot analysis of the expressed protein (Lane M: protein molecular weight markers; Lane 1: purified Cry11Aa). b Western blot analysis of the expressed protein (Lane M: protein molecular weight markers; Lane 1: purified biotinylated ATP-binding protein)
Fig. 5
Fig. 5
Bioassay of the ATP-binding protein. Each column represents the mean ± SEM (n ≥ 3). Each bar represents the mean ± SD of three technical replicates
Fig. 6
Fig. 6
Far-western blot analysis of Cry 11Aa and ATP-binding protein. a Far-western blot analysis (Lane M: protein molecular weight markers; Lane 1: purified Cry11Aa probed with biotinylated ATP-binding protein). b Far-western blot analysis (Lane M: protein molecular weight markers; Lane 1: purified ATP-binding protein probed with Cry11Aa)
Fig. 7
Fig. 7
ELISA analysis between Cry11Aa and ATP-binding protein. a Total binding of ABP to Cry11Aa and b total binding of Cry11Aa binding to ABP. Each bar represents the mean ± SD of three technical replicates
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