CMAH基因在小鼠不同组织中的转录水平分析  

Analysis of transcriptional levels of CMAH in different organ tissues of mice

作　　者：李一帆[1,2] 杨娜 王瑶[1,2] 尹德琦 赵旭[1,2] 桑晓宇 汉丽梅 

机构地区：[1]沈阳农业大学畜牧兽医学院,沈阳110866 [2]辽宁省人兽共患病研究重点实验室,沈阳110866

出　　处：《中国比较医学杂志》2018年第1期70-75,共6页Chinese Journal of Comparative Medicine

基　　金：校博士科研启动经费(编号:880416012)

摘　　要：目的哺乳动物细胞中所含唾液酸及其衍生物主要包括N-乙酰神经氨酸(N-acetylneuraminic acid,Neu5Ac)、N-羟乙酰神经氨酸(N-glycolylneuraminic acid,Neu5Gc)和脱氨神经氨酸(deaminoneuraminic acid,KDN),其中Neu5Ac可在胞苷单磷酸-N-乙酰神经氨酸羟化酶(cytidine monophospho-N-acetylneuraminic acid hydroxylase,CMAH)的催化下合成Neu5Gc。本研究应用相对荧光定量PCR方法检测BALB/c小鼠不同组织中CMAH基因的转录水平,为进一步分析不同组织中所含Neu5Gc含量差异提供参考依据。方法在NCBI数据库中查找CMAH基因mRNA序列并设计特异引物,选择小鼠β-actin为内参基因,应用相对荧光定量PCR方法检测正常BALB/c小鼠9种组织中CMAH基因mRNA的转录情况。结果在9种不同的小鼠组织中,肝脏组织中CMAH基因mRNA转录水平最高,其分别为脾脏的2.46倍、肾脏的3.17倍、气管的5.14倍、肺脏的11.70倍、心肌的21.12倍、骨骼肌的31.37倍、小肠的66.90倍以及脑组织的1055.99倍。结论本研究确定BALB/c小鼠CMAH基因在多种组织中均有转录且差异较大。Objective Sialic acid and its derivatives in mammalian cells mainly include N-acetylneuraminic acid（ Neu5 Ac）,N-glycolylneuraminic acid（ Neu5 Gc） and deaminoneuraminic acid（ KDN）,among which Neu5 Ac can be catalyzed by cytidine monophospho-N-acetylneuraminic acid hydroxylase（ CMAH） for the synthesis of Neu5 Gc. In this study,the transcriptional level of CMAH in different tissues of BALB/c mice were determined by relative fluorescence quantitative PCR,to provide a reference for further analysis of Neu5 Gc levels in different tissues. Methods The mRNA sequence of CMAH was searched in the NCBI database and specific primers were designed. The mouse β-actin was selected as an internal control,and the transcriptional levels of CMAH in 9 different organ tissues of BALB/c mice were detected by relative fluorescence quantitative PCR using SYBR Green dye. Results Among the 9 mouse organs,the transcriptional level of CMAH in the liver tissue was the highest,which was 2. 46 times higher of that in the spleen,3. 17 times of the kidney,5. 14 times of the trachea,11. 70 times of the lung,21. 12 times of the myocardium,31. 37 times of the skeletalmuscles,66. 90 times of the small intestine and 1056. 99 times of the brain tissue,respectively. Conclusions CMAH is transcribed in many organ tissues of mice,and its transcriptional levels vary in a quite wide range.

关 键 词：CMAH 唾液酸 相对荧光定量PCR BALB/C小鼠 

分 类 号：R-33[医药卫生]