测定猪不同组织基因表达时RT-PCR内参基因的选择

摘要

Taking different organizations of 95 kg weight fatten pigs,the expressions of 11 housekeeping genes (SDHA),(HMBS),(TOP2B),(ACTB),(B2M),(PPIA),(GAPDH),(HPRT1),(RPL4),(TBP) and (YWHAZ) were detected using real-time quantitative PCR.And the stability of internal genes was evaluated by geNorm program.Results of geNorm program analysis showed that TBP and RPL4 were stable expression internal genes in muscle tissue, GAPDH and RPL4 were stable expression internal genes in heart tissue, TBP and HPRT1 were stable expression internal genes in liver tissue, SDHA and ACTB were stable expression internal genes in spleen tissue, B2M and HPRT1 were stable expression internal genes in lung tissue.Thus it can be seen that the most stable expression internal genes in different tissues of pig are different,and the internal genes to standardize target genes in different tissues are also different.%以95 kg体重育肥猪的不同组织为研究对象,应用实时荧光定量PCR技术,对SDHA、HMBS、TOP2B、ACTB、B2M、PPIA、GAPDH、HPRT1、RPL4、TBP和YWHAZ共11个候选内参基因表达量进行检测,应用geNorm 程序对内参基因的稳定性进行分析.结果表明,肌肉组织中稳定表达的内参基因是TBP、RPL4,心脏组织中稳定表达的内参基因是GAPDH、RPL4,肝脏组织中稳定表达的内参基因是TBP、HPRT1,脾脏组织中稳定表达的内参基因是SDHA、ACTB,肺脏组织中稳定表达的内参基因是B2M、HPRT1,肾脏组织中稳定表达的内参基因是B2M、TBP.由此可见,不同组织中最稳定表达的内参基因不同,不同组织中标准化目的基因的内参基因也不一样.