摘要
内含子剪切是真核生物转录后修饰重要环节,是重要的基因表达调控方式。植物中多数基因存在可变剪切,能形成不同的剪切形式。然而对植物可变剪切因子的研究尚待深入。在本文的报告系统中,编码绿色荧光蛋白的基因中间插入了一个含有终止密码子的内含子以中断荧光蛋白的表达,而当内含子被正确剪切时能表达出绿色荧光蛋白。通过在本氏烟草(Nicotiana benthamiana)中进行瞬时表达实验,该系统可对植物可变剪切的效率进行定性与定量评估。此报告系统的建立为研究剪切辅助因子与特定前体mRNA的直接调控关系提供了新工具,同时也为快速筛选和研究剪切因子修饰对剪切过程的影响提供了便捷手段,有望进一步深化对可变剪切因子的认识。
Intron splicing is a crucial step in the post-transcriptional modification of eukaryotic organisms and serves as an important mean of gene expression regulation.Most genes in plants exhibit alternative splicing,resulting in different splicing forms.However,the study of splicing factors in plants remains to be furthered.Within this reporter system,gene encoding green fluorescent protein was inserted with a stop codon containing an intron to interrupt the expression of fluorescent protein unless it is properly spliced out.Through transient expression experiments in tobacco(Nicotiana benthamiana),this system can qualitatively and quantitatively evaluate the efficiency of alternative splicing in planta.The establishment of this reporter system provides a new tool for investigating the direct regulatory relationship between splicing factors and specific precursor mRNAs,as well as an expedient means for the rapid screening and study of the impact of splicing factor modifications during the splicing process.It is expected to further enhance our understanding of alternative splicing factors.
作者
唐茜
聂奕
王文慧
陈雨晴
魏济帆
黄予
陈怡君
刘柯颍
蔡文国
TANG Xi;NIE Yi;WANG Wenhui;CHEN Yuqing;WEI Jifan;HUANG Yui;CHEN Yijun;LIU Keying;CAI Wenguo(College of Life Science and Technology,Guangxi University,Nanning,530004;State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,Nanning,530004)
出处
《基因组学与应用生物学》
北大核心
2025年第5期443-454,共12页
Genomics and Applied Biology
基金
国家自然科学基金项目(32460085)
广西壮族自治区创新研究团队项目(2024GXNSFGA010003)
广西大学高层次人才启动经费项目(A3130051023)
亚热带农业生物资源保护与利用国家重点实验室青年学术骨干培养资助项目(SKLCUSAa202404)共同资助。
关键词
可变剪切
报告系统
荧光蛋白
本氏烟草
Alternative splicing
Reporter system
Fluorescent protein
Nicotiana benthamiana
