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journal1 ›› 2018, Vol. 34 ›› Issue (5): 686-692.DOI: 10.16409/j.cnki.2095-039x.2018.05.006

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Expression and Characteration of the Chitin Deacetylase 5 Gene from Holotrichia parallela Larvae

ZHAO Dan1, YAN Xiaoping1, LU Xiujun1, GUO Wei1,2   

  1. 1. College of Plant Protection, Agricultural University of Hebei, Baoding 071001, China;
    2. Plant Science and Technology College, Beijing University of Agriculture, Beijing 102206, China
  • Received:2018-05-18 Online:2018-10-08 Published:2018-10-15

Abstract: The HpCDA5 gene encoding chitin deacetylase was obtained by immune-screening the cDNA library of Holotrichia parallela with an anti-PM proteins polyclonal antiserum. The HpCDA5 protein has a polysaccharide deacetylase-like domain, belonging to Group V CDA. The HpCDA5 was successfully expressed in insect cells (sf9) as secreted proteins using recombinant baculoviruses (Bac to Bac system). Western blot analysis showed that the apparent molecular weight for recombinant HpCDA5 were about 42 kDa. qRT-PCR approach revealed that HpCDA5 has high expression in midgut. Subsequent chitin binding assays demonstrated that HpCDA5 had chitin binding affinity. The characterization of recombinant HpCDA5 laid a foundation for further studies and physiological functions of the HpCDA5 protein, and provide a theoretical basis for the biocontrol of H. parallela using CDA protein as target.

Key words: Holotrichia parallela, HpCDA5, insect cell expression, tissue expression, chitin binding affinity

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