Cloning and Expression of ABCB3 Gene in Beet Armyworm,Spodoptera exigua and Analysis of Its Binding Affinity with Bt Cry1Ab35

doi:10.16409/j.cnki.2095-039x.2023.01.052

Abstract

Abstract: The mechanism of Cry1Ab protein against Spodoptera exigua was studied in this study through analysis of the binding characteristics of ATP-binding cassette transporter with Bt Cry1Ab35 protein.A full-length cDNA sequence of SeABCB3(GenBank accession number OP948097) was cloned with a open reading frame of 3915 bp,which encodes 1304 amino acids.The SeABCB3 protein is predicted to have a molecular weight of 142.66 kDa.A SeABCB3-OM cDNA sequence coding for the extracellular region was further cloned and the prokaryotic expression vector pET30a-SeABCB3-OM was successfully constructed.Both Ligand blot and ELISA analysis indicated that SeABCB3-OM recombinant protein exhibited a specific binding to Bt Cry1Ab35 activated protein,indicating that it may be the receptor of Bt Cry1Ab35 protein.Quantitative Real-time PCR was used to analyze the expression level of SeABCB3 in different instar larvae,tissues,and structures.The results showed that SeABCB3 was expressed in all instar larvae and different tissues and structures of S.exigua,with the highest expression in the egg stage and the Malpighian tubule.In this study,a full-length cDNA of SeABCB3 in S.exigua was cloned,the SeABCB3-OM expression pattern was clarified in different instar larvae,tissues,and structures,and the binding characteristic of SeABCB3-OM protein to Bt Cry1Ab35 protein was determined,which paves a scientific basis for further exploration of the Bt Cry protein action mechanism against S.exigua.

Key words: Spodoptera exigua, ABC transporter, cloning and expression, expression analysis

CLC Number:

Q965

GUO Xiaochang, ZHAO Dan, WANG Qian, WANG Guandi, WEN Jingxin, WU Han, GUO Wei. Cloning and Expression of ABCB3 Gene in Beet Armyworm,Spodoptera exigua and Analysis of Its Binding Affinity with Bt Cry1Ab35[J]. Chinese Journal of Biological Control, 2024, 40(2): 327-336.

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