摘要
紫花苜蓿的秋眠与光敏色素基因的表达紧密相关。为了建立一种快速、灵敏检测紫花苜蓿PHYA和PHYB基因mRNA表达水平的实时逆转录聚合酶链反应(FQ-PCR)体系,本研究提取紫花苜蓿叶片总RNA,PCR扩增Actin、PHYA和PHYB基因片段,将其克隆入pMD18-T载体进行测序。基于SYBR Green I双链嵌合染料建立检测紫花苜蓿PHYA和PHYB基因mRNA表达水平方法,对PCR产物的熔解曲线进行分析评价其特异性,经测序鉴定目的片段已插入pMD18-T载体内,重组质粒的构建非常成功。为验证其准确性,我们检测紫花苜蓿WL-232 PHYA、PHYB mRNA表达量,结果证明:所建立的测定PHYA和PHYB基因mRNA水平的方法准确,适用于紫花苜蓿的各种组织的大量样本检测。
The fall dormancy of alfalfa is closely related with the expression levels of phytochrome genes.In order to establish a rapid and sensitive real-time reverse transcription-polymerase chain reaction(FQ-PCR) system for detecting the expression levels of PHYA and PHYB mRNA in alfalfa,total RNA was extracted from alfalfa leaves,and the partial sequences of Actin,PHYA and PHYB genes were cloned by RT-PCR.Subsequently,the method of measuring the expression levels of PHYA and PHYB mRNA in alfalfa was set up by ana...
出处
《江西农业学报》
CAS
2011年第4期96-100,共5页
Acta Agriculturae Jiangxi
基金
国家自然科学基金项目"光敏色素B调控苜蓿秋眠的分子机理研究"(30771527)
