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Regulation analysis of AcMNPV-mediated expression of a Chinese scorpion neurotoxin under the IE1, P10 and PH promoter in vivo and its use as a potential bio-insecticide

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Abstract

Objective

To analyze the regulation mechanism of AcMNPV (Autographa californica multicapsid nucleopolyhedrovirus)—mediated expression of BmK IT under IE1, P10 and PH promoters in the larva of Heliothis armigera..

Results

The transcription level of BmK IT gene in midgut and epidermal tissue was analyzed by quantitative PCR. The start time of transcription of recombinant BmK IT gene was early under the regulation of IE promoter, whereas transcription of BmK IT was high under the regulation of P10 promoter in the midgut tissue of infected larvae. TdT-UTP nick-end labeling (TUNEL) assay showed the degree of apoptotic cell death in the midgut tissue of AcMNPV-BmK IT-transfected insect larvae was higher than that in the AcMNPV treatment group at 8 h post-infection. The time–effect relationship between the insect’s humoral immunity and regulation of promoters was confirmed in the phenoloxidase activity assay.

Conclusion

The anti-insect mechanism and regulation of different promoters in AcMNPV-BmK IT at molecular and cellular levels provide an experimental basis for the development of recombinant baculovirus biopesticides.

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Acknowledgments

This project was supported by grants from ‘National Natural Science Foundation of China (Nos. 31272100, 31372199 and 31400765)’, ‘Natural Science Foundation of Shanxi Province (No. 2014011038-1)’, and ‘the Program for the Top Young Academic Leaders of Higher Learning Institutions of Shanxi’.

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Authors and Affiliations

  1. Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan, 030006, People’s Republic of China

    Yuejun Fu, Xing Li, Jun Du & Aihua Liang

  2. University of Miami Miller School of Medicine, Miami, FL, 33136, USA

    Shuhua Zheng

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  1. Yuejun Fu
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  2. Xing Li
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  3. Jun Du
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  4. Shuhua Zheng
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  5. Aihua Liang
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Correspondence to Yuejun Fu.

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Fu, Y., Li, X., Du, J. et al. Regulation analysis of AcMNPV-mediated expression of a Chinese scorpion neurotoxin under the IE1, P10 and PH promoter in vivo and its use as a potential bio-insecticide. Biotechnol Lett 37, 1929–1936 (2015). https://doi.org/10.1007/s10529-015-1875-4

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  • DOI: https://doi.org/10.1007/s10529-015-1875-4

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