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Characterization, cloning, and heterologous expression of a subtilisin-like serine protease gene VlPr1 from Verticillium lecanii

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Abstract

The entomopathogenic fungus Verticillium lecanii is a well-known biocontrol agent. V. lecanii produces subtilisin-like serine protease (Pr1), which is important in the biological control activity of some insect pests by degrading insect cuticles. In this study, a subtilisin-like serine protease gene VlPr1 was cloned from the fungus and the VlPr1 protein was expressed in Escherichia coli. The VlPr1 gene contains an open reading frame (ORF) interrupted by three short introns, and encodes a protein of 379 amino acids. Protein sequence analysis revealed high homology with subtilisin serine proteases. The molecular mass of the protease was 38 kDa, and the serine protease exhibited its maximal activity at 40°C and pH 9.0. Protease activity was also affected by Mg2+ and Ca2+ concentration. The protease showed inhibitory activity against several plant pathogens, especially towards Fusarium moniliforme.

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Authors and Affiliations

  1. College of Plant Sciences, Jilin University, Changchun, Jilin, 130062, P. R. China

    Gang Yu, Jin-Liang Liu, Li-Qin Xie, Xue-Liang Wang, Shi-Hong Zhang & Hong-Yu Pan

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  1. Gang Yu
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  2. Jin-Liang Liu
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  3. Li-Qin Xie
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  4. Xue-Liang Wang
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  5. Shi-Hong Zhang
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Correspondence to Hong-Yu Pan.

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Yu, G., Liu, JL., Xie, LQ. et al. Characterization, cloning, and heterologous expression of a subtilisin-like serine protease gene VlPr1 from Verticillium lecanii . J Microbiol. 50, 939–946 (2012). https://doi.org/10.1007/s12275-012-2199-x

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