Polyclonal Antibody Preparation, Eukaryotic Expression and Subcellular Localization of Frankliniella occidentalis FKBP6 Protein
WANG Xin1, ZHENG Li-Min2, ZHAO Xing-Yue3, MIAO Shu-Yue4, YE Qian2, ZHANG Yu2, SHI Xiao-Bin2, ZHANG De-Yong1,2,*, CHEN Jian-Bin2,*
1 Longping Branch, College of Biology, Hunan University, Changsha 410125, China; 2 Institute of Plant Protection, Hunan Academy of Agricultural Sciences, Changsha 410125, China; 3 College of Horticulture and Plant Protection, Inner Mongolia Agricultural University, Hohhot 010084, China; 4 Tobacco Research Institute, Henan Academy of Agricultural Sciences, Xuchang 461000, China
AbstractFrankliniella occidentalis is an invasive pest that harms global agricultural production. Exploring the biological functions of immunophilins FK506-binding protein 6 (FKBP6) of F. occidentalis can provide a new target for the prevention and control of F. occidentalis. In this study, FKBP6 (GenBank No. XP_026292958.1) of F. occidentalis was analyzed through bioinformatics, the sequence of FKBP6 gene (GenBank No. XM_026437173.1) was cloned, expression of FKBP6 protein was induced through prokaryotic cells, and the polyclonal antibody was prepared. Then, the protein was expressed by the Bac-to-Bac baculovirus system, the polyclonal antibody and its subcellular localization were detected through Western blot and immunofluorescence (IF) labeling. The amino acid sequence alignment and phylogenetic tree analysis showed that FKBP6 proteins among 20 insect species had conserved regions, but they were only highly related among species. FKBP6 protein in Sf9 cells was expressed through baculovirus systems, and then IF and Western blot dection showed that polyclonal antibodies specifically bound to FKBP6 protein. Furthermore, subcellular localization results showed that FKBP6 was localized in the cytoplasm. This study provides molecular conditions for the study of the biological function of FKBP6.
WANG Xin,ZHENG Li-Min,ZHAO Xing-Yue, et al. Polyclonal Antibody Preparation, Eukaryotic Expression and Subcellular Localization of Frankliniella occidentalis FKBP6 Protein[J]. 农业生物技术学报, 2024, 32(7): 1603-1613.
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