国家高技术研究发展计划(863)(2004AA2Z3170,2005AA2Z3D40),国家重点基础研究发展计划(2003CCA04300)和教育部重点项目(03007)资助.
This work was supported by grants from Hi-Tech Research and Development Program of China (2004AA2Z3170, 2005AA2Z3D40), National Basic Research Program of China (2003CCA04300) and Important Project of Ministry of Education of China (03007).
目前基因表达的转录分析多采用单一看家基因作为内参来校正目标基因的表达量. 实验中以人肝癌BEL-7402细胞为研究对象,应用实时荧光定量PCR技术,观察了新型三肽化合物酪丝缬肽作用后RPL13A、UBC、EIF4A、B2M、GAPDH和ACTB共6个看家基因mRNA水平的表达情况. 经过geNorm程序统计学分析处理,结果表明,这6个看家基因的表达存在差异,确定了RPL13A、UBC 2个看家基因用于校正目标基因的表达量. 基因表达转录分析中内参基因选择的必要性在实验中得以证明,更重要的是为各种实验因素影响下(尤其是新物质作用下)内参基因的选择介绍和提供了一种行之有效的方法.
At present, transcription analysis of gene expression commonly uses a single housekeeping gene as control for normalization. Through quantitative real-time PCR expression the levels of 6 housekeeping genes(including RPL13A,UBC,EIF4A,B2M,GAPDH,and ACTB) in BEL-7402 cell line were estimated under the role of a new tripeptide compound-YSV. Differences in expression levels were observed by analysis of geNorm program,and RPL13A, UBC were finally determined as suitable internal control genes. In conclusion, the necessity of choosing control genes was proved, and a good way was introduced to select control genes when experiments were handled by different empirical factors(especially under the effect of new materials).
张艳君,朱志峰,陆融,徐琼,石琳熙,简序,刘俊燕,姚智.基因表达转录分析中内参基因的选择[J].生物化学与生物物理进展,2007,34(5):546-550
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