应用半定量RT-PCR方法测定围产期奶牛肝PEPCK-C mRNA的丰度

Abundance of Liver PEPCK-C mRNA in Peripartum Dairy Cows Using Semi-quantity RT-PCR

建立了奶牛肝PEPCK-CmRNA丰度的半定量RT-PCR检测方法，其优化的PCR反应条件（25pL）为：57C退火、2．4mmol／L Mg^2＋、26～28个循环数、cDNA不小于2．6mg／L、2：1的PEPCK-C／β-actin引物比。同时，应用该方法检测了围产期奶牛肝中PEPCK-CmRNA丰度，结果显示：产前低血糖奶牛肝PEPCK-C mRNA丰度显著低于对照组，但是产后却高于对照组。围产期奶牛血糖水平与肝PEPCK-C mRNA丰度之间的相关系数，对照组为0．68（P=0．14），低血糖组为0．82（P=0．047）。因此，围产期奶牛肝PEPCK-CmRNA丰度对奶牛血糖浓度具有调节作用，低血糖奶牛表现得更为明显。

In the present study,a semi-quantity RT-PCR for detection of abundance of liver PEPCK-C mRNA in peripartum dairy cows was established. Its optimized parameters of PCR （25μL） were 57 C anneal, 2.4 mmol/L Mg^2＋, 26-28 cycle,minimum cDNA〉2.6 mg/L, 2 ： 1 primers ratio PEPCK-C to β-actin. Meanwhile,abundance of liver PEPCK-C mRN A in peripartum dairy cows was detected by this method. Results showed：In hypoglycemia dairy cows,the abundance of liver PEPCK-C mRNA at prepartum was lower than that in control,but at postpartum it was more than that in control. Correlation coefficiences between the blood sugar level and the liver PEPCK-C mRNA in peripartum dairy cows were 0.68（P=0. 04） in control cow and 0.82（P=0. 047） in hypoglycemia dairy cows. The results indicated that in peripartum dairy cows ,the abundance of liver PEPCK-C mRNA could regulate the blood sugar level,particurally in the dairy cows with hypoglycemia.