农业生物技术学报2024，Vol.32Issue(7)：1606-1613,8.DOI:10.3969/j.issn.1674-7968.2024.07.013
西花蓟马FKBP6蛋白多克隆抗体制备、真核表达及亚细胞定位
Polyclonal Antibody Preparation,Eukaryotic Expression and Subcellular Localization of Frankliniella occidentalis FKBP6 Protein
摘要
Abstract
Frankliniella 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.关键词
西花蓟马/FKBP6蛋白/抗体制备/Bac-to-Bac杆状病毒表达系统Key words
Frankliniella occidentalis/FKBP6 protein/Antibody preparation/Bac-to-Bac baculovirus expression system分类
农业科技引用本文复制引用
王鑫,郑立敏,赵星月,苗淑月,叶倩,张宇,史晓斌,张德咏,陈建斌..西花蓟马FKBP6蛋白多克隆抗体制备、真核表达及亚细胞定位[J].农业生物技术学报,2024,32(7):1606-1613,8.基金项目
国家重点研发计划政府间/港澳台重点专项(2018YFE0112600) （2018YFE0112600）
长沙市杰出创新青年培养计划(kq2009006) （kq2009006）
长沙市自然科学基金(kq2208140) （kq2208140）
国家自然科学基金(31871935 （）
32302300) （）
财政部和农业农村部:国家麻类产业技术体系资助(CARS-16-E17) （CARS-16-E17）
湖南省农业科学院科技创新项目(2023CX97) （2023CX97）
