Expression, Purification, and Characterization of Cuticular Protein Td14144 from Trypoxylus dichotomus

doi:10.16409/j.cnki.2095-039x.2022.03.005

Abstract

Abstract: Cuticular protein is an important component of insect cuticle. It plays a significant role in the growth and development of insects and may become the target for pest control. Trypoxylus dichotomus is an important pest of Dendrobium officinale, and there are few studies on its cuticular protein. The purpose of this article is to clarify the important functions of cuticular protein Td14144 from T. dichotomus by studying its expression, purification, and chitin-binding ability. Based on the transcriptome of T. dichotomus, the gene of Td14144 was cloned by RT-PCR (GenBank accession number: MZ463195) and subjected to bioinformatic analysis. Sequence analysis showed that Td14144 belongs to the cuticular protein of CPR_RR-2 family with the R&R domain. Phylogenetic analysis showed that Td14144 is closely related to AgCP of Anoplophora glabripennis. Subsequently, the gene was constructed into the pET-28a vector to generate the recombinant expression vector pET28a-14144. The vector was transformed into Escherichia coli BL21 for prokaryotic expression. Td14144 was purified by metal chelation chromatography and analyzed by SDS-PAGE and Western blot. The chitin binding ability of Td14144 toward different types of chitinous substrates including chitosan, α-chitin, β-chitin and colloidal chitin was evaluated. It was found that Td14144 showed the highest affinity to chitosan and α-chitin.

Key words: Trypoxylus dichotomus, cuticular protein, chitin- binding protein

CLC Number:

S476

LIU Jing, BAO Han, CHEN Lei, LIU Yuantao, LIU Tian, YANG Qing. Expression, Purification, and Characterization of Cuticular Protein Td14144 from Trypoxylus dichotomus[J]. Chinese Journal of Biological Control, 2022, 38(2): 513-520.

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