title: Characterization of a novel chitinase, DkChi, from Dendrolimus kikuchii nucleopolyhedrovirus.
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.
authors.affiliation: The Key Laboratory of Forest Protection, State Forestry Administration of China, Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, 2# Dongxiaofu, Haidian District, Beijing, 100091, People's Republic of China.
authors.forename: Qinghua
authors.initials: Q
authors.lastname: Wang
authors.forename: Liangjian
authors.initials: L
authors.lastname: Qu
authors.forename: Zhilin
authors.initials: Z
authors.lastname: Zhang
authors.forename: Yuzhu
authors.initials: Y
authors.lastname: Wang
authors.forename: Yongan
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D002688:Chitinases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D007306:Insecticides / Q000378:metabolism; D007915:Lepidoptera / Q000187:drug effects* / Q000821:virology*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017924:Nucleopolyhedroviruses / Q000201:enzymology* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D016019:Survival Analysis
chemical_list: D004279:DNA, Viral; D007306:Insecticides; D002688:Chitinases