Gene Cloning, Protein Expression and Chitin Binding Analysis of SeCDA8 from Beet armyworm, Spodoptera exigua

doi:10.16409/j.cnki.2095-039x.2022.01.017

Abstract

Abstract: Chitin deacetylase (CDA) catalyzes the deacetylation of chitin to chitosan, which plays an important role in the degradation of chitin. A full-length coding sequence (CDS) of SeCDA8 was obtained from the transcripome of Spodoptera exigua, named as SeCDA8 (GenBank number of OL689577) with an ORF of 1158 bp encoding 385 amino acid residues. Alignment of the sequence of SeCDA8 showed the highest sequence similarity to the sequence of CDA8 from Spodoptera litura by NCBI Blast and phylogenetic analysis. The prokaryotic recombinant expression vector pET30a-SeCDA8 was successfully constructed and transformed into E. coli Rosetta. The molecular weight of 45 kD recombinant protein was expressed in the precipitation after ultrasonic fragmentation. The chitin-binding activity of SeCDA8 protein to the regenerated chitin was higher than colloidal chitin. Both of them were weak. qRT-PCR analysis of SeCDA8 of midgut tissues from different regions showed that the SeCDA8 was more abundant both in the most anterior region of the midgut and the foregut, indicating that SeCDA8 may be related to the initial formation of peritrophic membrane. The results in this study provide an important theoretical basis for further exploring the physiological function of Group V CDA.

Key words: Spodoptera exigua, chitin deacetylase, expression, chitin-binding activity, tissue localization

CLC Number:

S188

WU Han, ZHAO Dan, GUO Xiaochang, LIU Zhaorui, GUO Wei, BI Yang. Gene Cloning, Protein Expression and Chitin Binding Analysis of SeCDA8 from Beet armyworm, Spodoptera exigua[J]. Chinese Journal of Biological Control, 2022, 38(6): 1508-1515.

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