中国马铃薯2025，Vol.39Issue(1)：1-7,7.DOI:10.19918/j.cnki.1672-3635.2025.01.001
马铃薯StBIN2蛋白的原核表达、酶活分析及多克隆抗体制备
Prokaryotic Expression,Enzyme Activity Analysis and Antibody Preparation of Potato StBIN2 Protein
摘要
Abstract
BIN2 is a serine/threonine protein kinase.Previous studies have revealed that BIN2 plays a crucial regulatory role in plant responses to both biotic and abiotic stresses.However,there is a lack of systematic research on the potato StBIN2 protein.To investigate the biological functions of the potato StBIN2 protein,the StBIN2 gene was obtained from potato via PCR amplification.The StBIN2 gene was then constructed into the pCold TF expression vector through homologous recombination to obtain the StBIN2 protein.The kinase activity of the StBIN2 protein was measured using a kinase activity kit.Additionally,purified StBIN2 protein was injected into New Zealand white rabbits to generate polyclonal antibodies.The full-length StBIN2 gene is 1 149 bp.The highest expression level of the StBIN2 protein was achieved under induction with 1.0 mmol/L isopropyl β-D-thiogalactoside(IPTG)at 16℃.The polyclonal antibodies prepared when the kinase activity of the StBIN2 protein was 1.09 U/L exhibited high specificity,making them suitable for Western blotting analysis.This study would lay a solid foundation for further elucidating the biological functions of the potato StBIN2 protein,contribute to a deeper understanding of the functional mechanisms of the StBIN2 gene in potato,and provide theoretical support and technical assistance for the genetic improvement and stress-resistant breeding of potato.关键词
马铃薯/BIN2蛋白/原核表达/多克隆抗体Key words
potato/BIN2 protein/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
王译晨,蒲文菲,彭锐,汪术艳,王居琴,刘石锋..马铃薯StBIN2蛋白的原核表达、酶活分析及多克隆抗体制备[J].中国马铃薯,2025,39(1):1-7,7.基金项目
四川省自然基金青年基金项目(2025ZNSFSC1104) （2025ZNSFSC1104）
西昌学院2025攀登计划人才引进专项项目(RCZ202506). （RCZ202506）
