猪TIGAR mRNA SYBR GreenⅠ荧光定量RT-PCR方法的建立  

Development of SYBR GreenⅠ-based RT-PCR Assay for Detection of TIGAR mRNA in Swine

作　　者：康大成 汤晓艳[1] 李朋颖 颜成英[2] 范成明[3] 

机构地区：[1]中国农业科学院农业质量标准与检测技术研究所,北京100081 [2]南京农业大学教育部肉品加工与质量控制重点实验室,江苏南京210095 [3]中国农业科学院作物科学研究所,北京100081

出　　处：《中国畜牧兽医》2014年第3期1-6,共6页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然基金(31000799)

摘　　要：根据GenBank中猪TP53诱导的糖酵解和凋亡调节因子(TP53-induced glycolysis and apoptosis regulator,TIGAR)mRNA序列(登录号:XM_001926543),设计合成了1对特异性引物,通过对反应条件的优化,建立了荧光定量RT-PCR技术检测猪TIGAR基因转录水平的方法,同时对长白猪宰后不同时间TIGAR mRNA转录水平进行了测定。结果表明,所建立的TIGAR基因标准曲线的R2为0.9995,内参基因β-actin标准曲线的R2为0.9996,2基因的扩增效率分别为103.7%和104.1%,扩增效率接近一致,可对TIGAR mRNA转录水平进行相对定量。该方法特异性强、灵敏度高,具有一定应用价值。According to swine TIGAR mRNA sequences available in GenBank （No：XM-001926543）, a pair of primers was designed, with experimental condition optimizing, the method of a Real-time fluorescent quantitative reverse transcription-poly merase chain reaction （RT PCR） was established to detect swine TIGAR gene, and the level of TIGAR mRNA of Landrace in different slaughter time was compared with the method established. The results showed that the R2 of standard curves of TI GAR and the housekeeper gene β- actin were 0. 9995 and 0. 9996, respectively, and the amplification efficiency of TIGAR （103.7%） was consistent with β-actin （104.1% ）. The methods established can be used for the determination of TIGAR mRNA in swine with high specificity and sensitivity. The research result was valuable to application.

关 键 词：猪 TIGAR MRNA SYBR Green Ⅰ RT-PCR 

分 类 号：S828[农业科学—畜牧学]