辐射条件下微藻基因表达内参基因的选择  

Selection of Suitable Internal Control Genes in Microalgae Under Radiation Condition

作　　者：蔡文凯[1,2] 胡金璐 李双双[1,2] 单革[3] 王高鸿[1] 

机构地区：[1]中国科学院水生生物研究所,武汉430072 [2]中国科学院大学,北京100049 [3]中国科学技术大学生命科学院,合肥230022

出　　处：《空间科学学报》2013年第6期651-658,共8页Chinese Journal of Space Science

基　　金：国家自然科学基金项目(30970688);载人航天项目共同资助

摘　　要：作为空间辐射的一种主要成分,紫外辐射可广泛引起陆地植物与水生生物细胞及其组份的破坏.荧光定量PCR技术广泛应用于各类胁迫环境下,研究目的基因的转录水平.在荧光定量PCR中选用合适的内参基因,能够更加准确地校正和标准化目的基因转录水平.本实验研究了辐射条件下水生生物莱茵衣藻6个传统内参基因18 S rRNA,GAPDH,β-actin,β-tubulin,EF1-α和UBC基因表达的稳定性.经GeNorm软件研究分析,在辐射条件下,莱茵衣藻18S rRNA基因表达最不稳定,而选用β-actin和GAPDH作为双内参,可以得到更精确的实验结果.As a major component of space radiation, ultraviolet radiation can extensively destruct terrestrial plants and aquatic. Real-time PCR technique has been widely used to detect the level of target mRNA expression when cell or tissues exposed to various types environmental stress. Choosing an appropriate internal control gene is important to accurately analyze the level of target gene transcription with real-time PCR technique. Therefore, our experiment tries to choose the most appropriate and stability reference gene from six traditional internal control genes, i.e., 18S rRNA, GAPDH,β-actin, β-tubulin, EF 1-α and UBC in the Chlamydomonas reinhardtii under radiation conditions. The results showed that C. reinhardtii 18S rRNA gene expression is the most unstable through GeNorm analysis, butβ-act;in and GAPDH gene were finally selected as a pair of suitable internal control genes in expression analysis, with which more accurate experimental results can be obtained.

关 键 词：辐射 莱茵衣藻 实时定量PCR 内参基因 

分 类 号：Q947.8[生物学—植物学] S917.3[农业科学—水产科学]