生物技术通报2024，Vol.40Issue(11)：192-201,10.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0423
山杏种子PsMATE40基因启动子的克隆及驱动表达分析
Cloning of the Promoter of MATE40 Gene from Prunus sibirica Seeds and Analysis of Gene Expression Driven by Promoter
摘要
Abstract
[Objective]The transporter protein PsMATE40 has been identified with an important role in the transport of seed amygdalin of Prunus sibirica.The aim of this study is to clone the specific promoter of PsMATE40 gene from P.sibirica seeds and to explore PsMATE40 expression driven by its native promoter,which should provide an important foundation for the application of PsMATE40 and its specific promoter.[Method]Based on our recent obtained sequence of PsMATE40 gene from the P.sibirica seeds,the full-length promoter fragment(designated as ProPsMATE40)of PsMATE40 gene was cloned by genome walking technique with genomic DNA of P.sibirica seeds as a template,and the cis-acting elements were identified within its promoter region by online programs.The GUS staining analysis was conducted to examine transcriptional activity of PsMATE40 gene promoter.Importantly,the plant expression vectors of PsMATE40 driven self-specific promoter(ProPsMATE40)and constitutive promoter(CaMV 35S)were constructed by the seamless cloning.Then it was transiently transformed into tobacco leaves by Agrobacterium-mediated method,and the transcriptional expression of PsMATE40 gene driven by them was comparatively analyzed.[Result]The full-length promoter sequence of PsMATE40 gene from P.sibirica seeds was 1 964 bp in length,containing two core promoter regulatory elements(CAAT-box and TATA-box)and several cis-acting regulatory elements in response to the hormone(auxin,jasmone acid and salicylic acid),stress(light,drought and wounding)and seed development,and importantly,the transcriptional level of PsMATE40 driven by its self-specific promoter(ProPsMATE40)was significantly higher compared with under the constitutive promoter(CaMV35S).[Conclusion]The expression of PsMATE40 gene from P.sibirica seeds is regulated by the complicated factors involved in plant hormones and biotic and abiotic stresses,and the self-specific promoter could effectively promote transcriptional expression of PsMATE40 gene.关键词
山杏种子/苦杏仁苷/PsMATE40基因/特异启动子/克隆/顺式调控元件/驱动表达分析Key words
Prunus sibirica seed/amygdalin/PsMATE40 gene/specific promoter/cloning/cis-acting regulatory elements/promoter-driven expression analysis引用本文复制引用
王紫睿,刘潇菡,修宇,林善枝..山杏种子PsMATE40基因启动子的克隆及驱动表达分析[J].生物技术通报,2024,40(11):192-201,10.基金项目
"十四五"国家重点研发计划(2022YFD2200400),国家自然科学基金面上项目(31972592) （2022YFD2200400）
