Expression, purification and characterization of galectin AcGalectin in Apis cerana cerana (Hymenoptera: Apidae)
YUE Jin-Jin1,#, MA Yu-Chi1,#, Liu Quan2, JIANG Li-Li1, FEI Dong-Liang1, ZHANG He1, SUN Li1, LI Ming1, MA Ming-Xiao1,*
(1. College of Animal Husbandry and Veterinary, Jinzhou Medical University, Jinzhou, Liaoning 121000, China; 2. Liaoning Province Animal Health Monitoring and Earlier Warning Center, Shenyang 110001, China)
Abstract 【Aim】 This study aims to explore the function of galectin in Apis cerana cerana and its interaction with the honeybee virus. 【Methods】 Galectin gene was amplified from A. cerana ceranas by RT-PCR using the extracted total RNA as the template, and then the sequence and structural characteristics of deduced amino acids were analyzed by using bioinformatics tools. The codons of the galectin gene were optimized based on the codon bias of Escherichia coli, and the recombinant protein was produced in E. coli and purified by affinity chromatography. The binding activities of the recombinant protein with Chinese sacbrood virus (CSBV), Chronic bee paralysis virus (CBPV) and Deformed wing virus (DWV) were analyzed by far-western blotting. 【Results】 A galectin gene was cloned from A. cerana cerana and named AcGalectin (GenBank accession no: MG557559), and its full-length cDNA is 1 473 bp in length. AcGalectin has a stable spatial structure containing a Gal-bind_lectin domain. The phylogenetic tree constructed based on the amino acid sequences from AcGalectin and the previously reported galectins indicated that galectins form two clusters, one related to galectins from the Hymenoptera, and the other related to galectins from other insects. The codon-optimized AcGalectin gene could be highly expressed in host bacteria BL21 (DE3), and the purified recombinant AcGalectin protein could bind with CSBV and CBPV, but not with DWV. 【Conclusion】 The results indicate that AcGalectin can bind with CSBV and CBPV, and this lays a foundation for the further study on the function of AcGalectin in the process of CSBV and CBPV infections.
YUE Jin-Jin,MA Yu-Chi,LIU Quan et al. Expression, purification and characterization of galectin AcGalectin in Apis cerana cerana (Hymenoptera: Apidae)[J]. ACTA ENTOMOLOGICA SINICA, 2018, 61(5): 546-554.