Abstract
Dendrolimus kikuchii Matsumura nucleopolyhedrovirus (DkNPV) is a novel nucleopolyhedrovirus strain that has exhibited high potential as biological control agent against D. kikuchii. In this work, a 1755-bp DkChi gene with sequence homology to a chitinase gene was cloned from the genomic DNA of DkNPV using a DNA fragment library. The DkChi gene, encoding 558 residues protein with a predicted mass of 61.6 kDa, was expressed at high levels in Escherichia coli and purified by affinity chromatography. We confirmed that the prepared protein was the DkChi protein by mass spectrometry analysis. Enzyme activity analysis showed that DkChi had both endo- and exo-chitinase activities. Interestingly, the DkChi protein displayed a strong insecticidal activity against Spodoptera exigua, Hyphantria cunea, Helicoverpa armigera and Lymantria dispar. The results suggest that DkChi is a good candidate protein for significantly contributing to pest control.
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Acknowledgments
This work was supported by the Program of National Nature Science Foundation of China (grant no. 31200491) and State 863 Project funded by Ministry of Science and Technology (grant no. 2012AA101503), P.R. China.
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705_2013_1775_MOESM1_ESM.tif
Fig. S1 Phylogenetic tree of chitinases from baculovirus. The tree was constructed by the neighbor-joining method, and bootstrap values are indicated at the branches. The amino acid sequences of chitinases were from the following viruses: Dendrolimus kikuchii NPV (JN680874), Hyphantria cunea NPV (YP473218), Orgyia pseudotsugata MNPV (NP046280), Bombyx mori NPV (NP047523), Autographa californica MNPV (NP054156), Spodoptera exigua MNPV (NP037779), Helicoverpa armigera SNPV (AAK28347) and Spodoptera litura NPV (NP258310) (TIFF 292 kb)
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Wang, Q., Qu, L., Zhang, Z. et al. Characterization of a novel chitinase, DkChi, from Dendrolimus kikuchii nucleopolyhedrovirus. Arch Virol 158, 2523–2530 (2013). https://doi.org/10.1007/s00705-013-1775-7
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DOI: https://doi.org/10.1007/s00705-013-1775-7