低表达甘蓝花药发育相关基因小体积荧光定量反应体系的建立和可靠性分析  

A Real-time Quantitative PCR Protocol with Small Reaction Volume for Studies of Low Expression Genes Related to Cabbage Anther Development

作　　者：王效维[1,2] 刘海霞[2] 杨颖丽[1] 康俊根[2] 

机构地区：[1]西北师范大学,兰州730070 [2]北京市农林科学院蔬菜研究中心,北京100097

出　　处：《西北农业学报》2011年第4期111-115,共5页Acta Agriculturae Boreali-occidentalis Sinica

基　　金："十一五"国家科技支撑计划项目(2009BADB8B03);北京市自然科学基金项目(6102012)

摘　　要：植物发育过程中很多重要基因拷贝数较低。为建立稳定可靠、适于低拷贝微量模板表达分析的实时qRT-PCR小体积反应体系,采用在甘蓝花药中表达量较少的AT-HOOK基因进行小反应体积荧光定量表达分析,研究小反应体积的扩增效率和可行性,并通过对其在甘蓝不同植物器官、不同花药组织和花药发育不同时期的表达特征的重复性和可靠性分析,最终建立了稳定可靠而且成本较低的10μL荧光定量分析体系,为进一步大规模深入研究甘蓝花药发育相关基因表达特征及发育调控基因功能分析奠定基础,同时也为其他低拷贝表达基因的研究提供参考。Many important genes have a relatively low expression level during plant development.To establish a cheap and reliable quantitative PCR protocol for expression studies of low-copy-number genes,we used the low expression gene AT-HOOK in cabbage anther to evaluate the feasibility of real-time quantitative PCR with small volume regarding the stability and efficiency.A feasible 10 μL real-time quantitative PCR protocol was established.The experiment here showed that the small volume quantitative PCR system was stable and reliable and it could be helpful for the extensive expression studies of low-copy-number genes related to cabbage anther development.

关 键 词：甘蓝 基因表达 小体积反应体系 实时荧光定量PCR 可行性 

分 类 号：R450[医药卫生—治疗学]