中国卤虫actin基因的实时荧光定量PCR方法的建立和优化  

Establishment and Improvement of Real-time Fluorescence Quantitative PCR for actin Gene of Artemia sinica

作　　者：侯林[1] 姜丽娟[1] 孙文静[1] 张瑞锋[1] 王家庆[1] 赵欣涛[1] 安家璐[1] 

机构地区：[1]辽宁师范大学生命科学学院,辽宁大连116029

出　　处：《辽宁师范大学学报（自然科学版）》2006年第4期466-469,共4页Journal of Liaoning Normal University:Natural Science Edition

基　　金：国家自然科学基金资助项目(30271035)

摘　　要：根据GenBank中卤虫actin基因的保守区域序列设计并合成一对引物,采用SYBR GreenⅠ作为染料建立了实时荧光定量PCR(real-time fluorescence quantitative PCR)方法.以Ct值为纵坐标,以稀释倍数的对数为横坐标,建立标准曲线,其相关系数达到了0.999.溶解曲线分析显示产物为单一的特异峰,Tm为83.6℃.结果表明:本实验建立的中国卤虫actin基因的实时荧光定量PCR法扩增效率高、特异性强、线性范围广、检测周期短,为actin基因作为内参基因进行卤虫早期胚胎发育基因表达的定量分析奠定了基础.According to the actin gene sequences of Artemia sinica available in GeneBank, a pair of primers was designed for establishing a SYBR Green Ⅰ quantitative real-time fluorescence quantitative PCR method. Standard curve was established basing on Ct vs initial input amount of Log （copy number）, and the correlation coefficient of it was 0. 999. The melting curve showed a single peak with a Tm of 83.6℃. The results demonstrated that method developed in this study can quickly detect actin in broad range with high efficiency, specificity. The experiment provided the basis for actin gene of Artemia sinica as a control gene in quantitative analysis of early embryonic development gene expression of Artemia sinica.

关 键 词：中国卤虫 ACTIN基因 实时荧光定量PCR 

分 类 号：Q95-336[生物学—动物学] Q959.223.12