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.