摘要
Web MicroRNA Designer(WMD3)是一个专门应用于植物人工miRNA设计和前体序列设计的专业网站,其中利用重叠PCR法合成amiRNA前体序列,涉及多个PCR和纯化回收过程,步骤繁琐,周期较长。本研究利用WMD3设计了麻竹转录因子DlSCL6基因的人工miRNA序列和其前体序列,采用改进的重叠PCR方法,快速克隆获得了该前体序列,并以改造的pCMABIA1301载体为框架,构建了该前体的植物表达载体,为进一步研究利用amiRNA调控SCL6基因的表达奠定了基础。
Web MicroRNA Designer（ WMD3） is a professional website specially used for sequence design of plant artificial miRNA and its precursor. Overlap PCR method was used for the synthesis of amiRNA precursor sequence,which involved multiple PCR purifications and recovery processes with complicated steps and a longer period. In this study,sequences of plant artificial miRNA and its precursor for DlSCL6 in Dendrocalamus latiflorus were designed according to WMD3 and an improved method of overlapping PCR was used for quickly cloning of the precursor sequence. Based on the frame of p CMABIA1301 vector,the expression vector of amiRNA precursor for Dl SCL6 was constructed,which laid a foundation for further study in the expression regulation of SCL6 using amiRNA.
出处
《世界竹藤通讯》
2016年第2期1-5,共5页
World Bamboo and Rattan
基金
国家林业公益性行业科研专项经费项目(编号:201504106)
"十二五"国家科技支撑计划专题(编号:2012BAD23B0504)
关键词
麻竹
人工miRNA
重叠PCR
表达载体构建
Dendrocalamus latiflorus
amiRNA
overlap PCR
construction of expression vector
