测定猪不同组织基因表达时RT-PCR内参基因的选择

发布时间：2018-03-18 00:08

本文选题：猪　切入点：实时荧光定量PCR　出处：《家畜生态学报》2017年06期 　论文类型：期刊论文

【摘要】：以95kg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。
[Abstract]:Using 95 kg body weight fattening pigs in different tissues as the research object, using real-time fluorescence quantitative PCR technique, the expression of 11 candidate internal reference genes of SDHAHAHMBSTOP2BACTBP2MGAPDHHPP-HPRT1HPL4TBP and YWHAZ were detected, and the stability of the internal reference genes was analyzed by geNorm program, the results showed that: 1. The stably expressed internal reference gene in muscle tissue is TBPnRPL4, the stable expression gene in heart tissue is GAPDHnRPL4, the stable expression gene in liver tissue is TBPnHPRT1, the stable expression gene in spleen tissue is SDHAACTB, and the stable gene in lung tissue is stably expressed in lung tissue. The stable expression of B2MN HPRT1 gene in renal tissue is B2MN TBP1, so it can be seen that the stable expression of B2MN HPRT1 gene in renal tissue is the same as that of B2MN TBP. The most stably expressed endogenous reference genes are different in different tissues, and the standard target genes are different in different tissues.
【作者单位】：青岛农业大学动物科技学院;山东省农业科学院畜牧兽医研究所;山东省畜禽疫病防治与繁育重点实验室;
【基金】：国家自然科学基金(31501928) 山东省科技发展计划(2015GNC111011) 山东省农业科学院青年基金(2014QNM41);山东省农业科学院科技创新重点项目(2014CX202) 山东省自然基金(ZR2015CM007)
【分类号】：S828

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