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AcMNPV-mediated expression of BmK IT promotes the apoptosis of Sf9 cells and replication of AcMNPV

FU Yue-Jun, ZHAO Jie, LIANG Ai-Hua, HU Feng-Yun

1Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, China； 2Shanxi Provincial People’s Hospital, Affiliate of Shanxi Medical University, Taiyuan 030012, China

Abstract

Chinese scorpion Buthus martensii Karsch (BmK) venom is a rich source of neurotoxins which bind to various ion channels with high affinity and specificity and thus widely used as compounds to modulate channel gating or channel currents. To promote the insecticidal effects of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV), the gene encoding an excitatory insect toxin, BmK IT, was inserted into the genome of AcMNPV to construct a recombinant baculovirus, AcMNPV-BmK IT. Spodopter frugiperda 9 (Sf9) cells were infected with AcMNPV and AcMNPV-BmK IT respectively for 24 h. Results from the MTT assay, TUNEL assay, analysis of the expression level of apoptosis-related proteins (c-Myc, cleaved-Caspase3, Bcl-2 and Bax) of Sf9 cells, the transcription level of key genes (38K, C42, P78, F) of AcMNPV, and viral propagation assay demonstrated that AcMNPV-mediated expression of BmK IT promoted the apoptosis of Sf9 cells and replication of AcMNPV. The results laid a foundation for further structural and functional analysis of BmK IT.

Key words: Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV); Buthus martensii Karsch insect toxin (BmK IT); Spodopter frugiperda 9 cell; apoptosis; replication

Received: 2014-05-19　　Accepted: 2014-11-04

Corresponding author: 付月君,胡风云　　E-mail: yjfu@sxu.edu.cn, fengyun71@163.com

Citing This Article：

FU Yue-Jun, ZHAO Jie, LIANG Ai-Hua, HU Feng-Yun. AcMNPV-mediated expression of BmK IT promotes the apoptosis of Sf9 cells and replication of AcMNPV. Acta Physiol Sin 2015; 67 (3): 305-311 (in Chinese with English abstract).