生物技术通报2024，Vol.40Issue(5)：66-73,8.DOI:10.13560/j.cnki.biotech.bull.1985.2023-1178
马铃薯线粒体靶向表达载体的构建与应用
Construction and Application of Potato Mitochondrial Targeted Expression Vector
摘要
Abstract
[Objective]The genetic variations of mitochondria genome usually lead to cytoplasmic male sterility(CMS)in plants.Biotechnology is an important measure to produce variations resulting in CMS lines.In order to establish an efficient expression vector system for the research on potato mitochondrial genes,the optimal potato mitochondrial targeting expression vector was explored by comparing the expression level of eGFP,which was promoted by different combinations of promoters and mitochondrial targeting signal peptides(MTS).[Method]Three promoters AtUBI10,StUBI10,2×35S and two mitochondrial transport signal peptides ATPγ and Rf1b were tested.The expression level of the target vector was verified by tobacco leaf transient expression system and PEG-mediated potato protoplast transformation system.[Result]AtUBI10::ATPγ-eGFP among 6 expressing vectors had the best expression effect in tobacco and was accurately located in mitochondria.The second best ones were StUBI10::ATPγ-eGFP and 2×35S::ATPγ-eGFP.The vector AtUBI10::ATPγ-eGFP also showed the best performance,followed by StUBI10::Rf1b-eGFP and AtUBI10::Rf1b-eGFP in the potato protoplast transformation system.The two vectors with 2×35S promoter were weakly expressed in protoplasts.[Conclusion]The comparative studies showed the mitochondrial vector constructed by the promoter AtUBI10 and the mitochondrial transport signal peptide ATPγ is the best combination,and this vectors will be of great benefit to the relative study on mitochondrial genes.关键词
马铃薯/线粒体转运信号肽/靶向表达载体/亚细胞定位/细胞质雄性不育Key words
potato/mitochondrial transport signal peptides/targeted expression vector/subcellular localization/cytoplasmic male sterility引用本文复制引用
张震,李清,徐菁,陈凯园,张春芝,祝光涛..马铃薯线粒体靶向表达载体的构建与应用[J].生物技术通报,2024,40(5):66-73,8.基金项目
项目基金:国家重点研发计划(2022YFF1002500),国家自然科学基金项目(32272725) （2022YFF1002500）
