利用实时荧光定量RT-PCR检测鸡A-FABP和H-FABP基因的差异表达  

Relative Expression of A-FABP and H-FABP Gens in Chicken Measured by Real-Time PCR

作　　者：屠云洁[1,2,3] 苏一军[2,3] 王克华[2] 张学余[2,3] 陈国宏[1] 

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009 [2]中国农业科学院家禽研究所,江苏扬州225003 [3]国家地方禽种资源基因库,江苏扬州225003

出　　处：《中国畜牧杂志》2010年第7期1-4,共4页Chinese Journal of Animal Science

基　　金：国家科技支撑计划(2008BADC1B03;2008BADC1B01;2006BDA01A09);国家863计划(2008AA101009-7)

摘　　要：依据鸡A-FABP和H-FABP基因及参照基因GAPDH序列设计引物,以鸡心肌、胸肌和腿肌、肝脏和腹脂总RNA逆转录合成的cDNA为模板,GAPDH基因为参照基因,应用SYBRGreenI实时荧光定量PCR检测如皋黄鸡和安卡红鸡A-FABP和H-FABP基因的差异表达。结果表明:A-FABP、H-FABP基因和参照基因GAPDH基因融解曲线为单峰,无杂峰及二聚体,其扩增效率分别为99.7%、99.8%和100.0%。如皋黄鸡H-FABP基因在心肌、胸肌和腿肌的差异表达量分别是安卡红鸡的0.0860、0.0680倍和0.0580倍(P<0.05)。H-FABP基因mRNA表达量与肌内脂肪含量呈显著负相关。如皋黄鸡A-FABP基因在腹脂和肝脏的表达量分别是安卡红鸡的15.9640倍和10.9640倍(P<0.05),而在心肌、胸肌和腿肌的表达量差异不显著。The relative expression in heart and adipocyte fatty acid-binding protein （H-FABP and A-FABP ） genes was detected using real time quantitative RT-PCR in Rugao and Anka chicken. The primers were designed according to the sequences of H-FABP, A-FABP and GAPDH gene. GAPDH gene was the internal reference gene. The cDNA fragment was amplified from leg muscle, breast muscle, cardiac muscle, liver and abdominal fat mRNA; H-FABP, A-FABP and GA PDH gene expression was detected using SYBR Green I real-time fluorescent quantity PCR. Melting curve analysis showed a single peak of H-FA BP, A-FABP gene and GA PDH gene. PCR efficiency of H-FA BP, A-FABP and GA PDH was 99.7%, 99.8% and 100.0% respectively. The relative H-FABP mRNA level （ratio of Anka to Rugao） in cardiac muscle was 0.0860, 0.0680 and 0.0580, significantly higher than that of the other tissues （P 〈 0.01）. H-FABP mRNA expression level was significantly negative correlation with IMF contents. The relative A -FABP mRNA level （ratio of Anka toRugao） in abdomen fat and liver was 15.9640 and 10.9640, significantly higher than that of the other tissues （P 〈 0.01）. The relative A -FABP mRNA level in abdomen fat was higher than that of Liver. There were no significant differences between leg muscle, breast muscle and cardiac muscle of relative A -FA BP mRNA levels.

关 键 词：荧光定量逆转录PCR A-FABP基因 H-FABP基因 相对定量 鸡 

分 类 号：S831.2[农业科学—畜牧学]