摘要
目的:研究microRNA169b(miRNA169b)启动子不同长度片段的活性及自身调控。方法:用生物信息学方法分析水稻中miRNA169b前体上游1500 bp序列,并从水稻幼苗基因组DNA中扩增miRNA169b前体上游500、1000和1500 bp启动子片段,分别将3个片段克隆至萤光素酶报告基因载体p5XGal4-LUC上。结果:构建了miRNA169b启动子重组载体,序列分析表明与预期结果一致。结论:miRNA169b启动子调控的萤光素酶报告基因表达载体的构建,为研究水稻中miRNA基因自身的转录奠定了基础。
Objective:To research activity and self-regulation of different segments of miRNA169b promoter in rice.Methods:The 1500,1000 and 500 bp DNA segment of rice miRNA169b promoter were analyzed with bioinformatics and amplified by PCR from rice seedlings and cloned to p5XGal4-LUC vectors,then the positive clones were confirmed by restriction enzymes and sequencing.Results:The targeted segments were cloned to Firefly luciferase report vectors p5XGal4-LUC.Conclusion:These results may make an important basis for future studies of miRNA gene promoter activity and regulation of gene expression.
出处
《生物技术通讯》
CAS
2011年第2期234-237,共4页
Letters in Biotechnology
基金
国家自然科学基金(30971704)