At present, transcription analysis of gene expression commonly uses housekeeping genes as control for normalization. In this study, the expression levels of three housekeeping genes including GAPDH, β-actin, and 18S ...At present, transcription analysis of gene expression commonly uses housekeeping genes as control for normalization. In this study, the expression levels of three housekeeping genes including GAPDH, β-actin, and 18S rRNA in six tissues and five developmental stages of the Mandarin fish Siniperca chuatsi were assayed with quantitative real-time PCR （qPCR）. Differences in expression levels were analyzed using geNorm program. The results demonstrate that β-actin is the most stable gene at developmental stages and GAPDH is the most stable in different tissues. While 18S rRNA expression during development is differentially regulated, which indicates it is suitable as an internal control for gene expression normalization at the developmental level. Overall, the data suggest that the two most stable housekeeping genes are enough to accurately calibrate gene expression in S. chuatsi. The significance of this study provided convincing references and methodology for housekeeping gene selection and normalization in gene expression analysis with regular PCR or qPCR.展开更多
苹果果实着色期基因表达水平的变化对果实品质形成具有重要影响,选择适合的内参基因是提高实时荧光定量PCR分析准确性的首要条件。本试验以‘富士’苹果着色过程中不同取样时间的果皮为材料,通过q RT-PCR分析了常用候选持家基因β-actin...苹果果实着色期基因表达水平的变化对果实品质形成具有重要影响,选择适合的内参基因是提高实时荧光定量PCR分析准确性的首要条件。本试验以‘富士’苹果着色过程中不同取样时间的果皮为材料,通过q RT-PCR分析了常用候选持家基因β-actin、EF-1α、GAPDH和18S r RNA的表达变化,借助ge Norm和Norm Finder程序筛选出在果实着色期q RT-PCR分析的理想内参基因。结果表明,EF-1α的表达水平高且最稳定,其次是18S r RNA,而β-actin和GAPDH的相对表达水平较低;同时,用筛选的内参基因EF-1α和18S r RNA分析苹果花青苷合成途径的二氢黄酮醇-4-还原酶基因的表达水平,其表达规律比较一致,均呈现正态分布。因此,EF-1α和18S r RNA是研究苹果着色期表达的理想的内参基因。展开更多
基金国家自然科学基金(3077164430972263)Aid Program for Science and Technology Innovative Research Team in Higher Educational Instituions of Hunan Province
文摘At present, transcription analysis of gene expression commonly uses housekeeping genes as control for normalization. In this study, the expression levels of three housekeeping genes including GAPDH, β-actin, and 18S rRNA in six tissues and five developmental stages of the Mandarin fish Siniperca chuatsi were assayed with quantitative real-time PCR （qPCR）. Differences in expression levels were analyzed using geNorm program. The results demonstrate that β-actin is the most stable gene at developmental stages and GAPDH is the most stable in different tissues. While 18S rRNA expression during development is differentially regulated, which indicates it is suitable as an internal control for gene expression normalization at the developmental level. Overall, the data suggest that the two most stable housekeeping genes are enough to accurately calibrate gene expression in S. chuatsi. The significance of this study provided convincing references and methodology for housekeeping gene selection and normalization in gene expression analysis with regular PCR or qPCR.
文摘苹果果实着色期基因表达水平的变化对果实品质形成具有重要影响,选择适合的内参基因是提高实时荧光定量PCR分析准确性的首要条件。本试验以‘富士’苹果着色过程中不同取样时间的果皮为材料,通过q RT-PCR分析了常用候选持家基因β-actin、EF-1α、GAPDH和18S r RNA的表达变化,借助ge Norm和Norm Finder程序筛选出在果实着色期q RT-PCR分析的理想内参基因。结果表明,EF-1α的表达水平高且最稳定,其次是18S r RNA,而β-actin和GAPDH的相对表达水平较低;同时,用筛选的内参基因EF-1α和18S r RNA分析苹果花青苷合成途径的二氢黄酮醇-4-还原酶基因的表达水平,其表达规律比较一致,均呈现正态分布。因此,EF-1α和18S r RNA是研究苹果着色期表达的理想的内参基因。
