中国蔬菜Issue(9)：64-73,10.DOI:10.19928/j.cnki.1000-6346.2025.3005
马铃薯StDGKs的逆境响应及蛋白表达纯化研究
Study on Stress Response and Protein Expression Purification of Potato StDGKs
摘要
Abstract
Potato(Solanum tuberosum L.),the fourth-largest staple food crop globally,requires enhanced genetic improvement through the identification of stress-resistance genes.Plant diacylglycerol kinase(DGK)play a crucial role in stress responses by regulating the diacylglycerol(DAG)-phosphatidic acid(PA)signaling pathway.However,the functional mechanisms of potato DGK family members(StDGKs)under abiotic stress remain insufficiently characterized.In this study,we performed comprehensive bioinformatics analyses to examine the amino acid sequences and protein domains of StDGKs.Using quantitative real-time PCR(qRT-PCR),we assessed their expression profiles under drought,salt,and low-temperature stress conditions.Furthermore,we achieved prokaryotic expression and Ni-NTA affinity purification of six StDGKs using the E.coli BL21(DE3)system.Our findings demonstrate that StDGKs gene expression is significantly induced by abiotic stressors,with high conservation observed in their protein sequences and structural domains.All six recombinant StDGK proteins were successfully expressed as NusA-His-tagged fusion proteins at 37℃ under 1.0 mmol·L-1 IPTG induction,and affinity chromatography yielded purified products with the expected molecular weights.This study confirms that StDGKs are responsive to abiotic stress and that their encoded proteins can be efficiently expressed and purified in vitro.These results provide valuable candidate gene resources for elucidating stress-resistance mechanisms and advancing the development of stress-tolerant potato cultivars.关键词
马铃薯/二酰基甘油激酶/非生物胁迫/原核表达/蛋白质纯化Key words
potato/diacylglycerol kinase/abiotic stress/prokaryotic expression/protein purification引用本文复制引用
曾相琴,王迪,夏天,黄彦,田周娥,李萌,李振,朱秀春,林原..马铃薯StDGKs的逆境响应及蛋白表达纯化研究[J].中国蔬菜,2025,(9):64-73,10.基金项目
国家自然科学基金项目(31901586),湖南省研究生科技创新项目(CX20220723) （31901586）
