【作者】 刘丽； 蒋国民； 李金龙； 陈湘艺； 王晓清； 李传武；

【Author】 Liu Li;Li Chuanwu;Peng Zhitao;Deng Shiming;Li Jinlong;Wang Xiaoqing;Chu Wuying;College of Animal Science and Technology, Hunan Agriculture University;Fisheries Research Institute of Hunan Province;

【机构】 湖南农业大学动物科学技术学院； 湖南省水产科学研究所；

【摘要】 RT-qPCR定量分析中,通常采用内参基因来校正目标基因的表达量,从而获得精确、可靠的实验结果。本研究选用GAPDH、18S rRNA、RPL13、RPL19、RPS2和β-actin 6个内参基因,利用Norm Finder、ge Norm与Bestkeeper三个软件,进行筛选和优化中华鳖两个组织中表达最稳定的内参基因。结果发现,六个内参基因中,RPL13在中华鳖肌肉组织中表达最稳定,而RPL19在肝脏中表达最稳定。因此,RPL13和RPL19适用于校正目标基因的表达量,此结果为中华鳖肌肉和肝脏组织中目标基因的表达奠定了坚实基础。更多还原

【Abstract】 The reference gene is used to calibrate the expression level of the target gene to obtain accurate and reliable result in RT-qPCR analysis. In our study, the optimal reference genes were detected and chosen from six candidate reference genes including GAPDH, 18 S rRNA, RPL13,RPL19, RPS2 and β-actin which were analyzed in q RT-PCR method by using Ge Norm,Norm Finder and Bestkeeper softwares. The resulted indicated that the most stable reference genes was RPL13 in the skeletal muscle, but the most stable reference genes was RPL19 in the liver among 6 reference genes. In conclusion, RPL13 and RPL19 were used to normalize m RNA levels of target genes in the muscle and liver tissues of Chinese softshell turtle. The result laid a sound foundation for studying of the expression levels of target genes.更多还原