黑羽番鸭LPL基因克隆及在肌肉发育中的表达规律

Cloning of LPL gene and its mRNA expression regulation in muscle tissue of black Muscovy duck

脂蛋白脂酶（LPL）是水解甘油三酯的关键性酶,可以为细胞提供游离的甘油三酯,并作用于脂蛋白循环,与机体的脂质代谢有密切关系。本研究利用同源克隆策略克隆黑羽番鸭LPL基因,并采用实时荧光定量PCR技术分析其在肌肉组织发育过程中mRNA表达规律。结果表明,黑羽番鸭LPL基因cDNA序列长度为1515 bp,包括了1485 bp的编码区,编码494个氨基酸。与禽类LPL基因核苷酸和氨基酸序列同源性均在90%以上,与人和哺乳动物同源性在70%~75%之间。实时荧光定量PCR检测结果显示,LPL基因在第2周龄胸肌中mRNA表达量较低,然后开始上升,达到最大值再出现下降并保持在一定水平上;公鸭腿肌表达规律和公鸭一致,但是母鸭腿肌表达量呈波浪状。

Lipoprotein lipase is a key enzyme in development of lipid metabolism that acts on hydrolyzing triglycerides, providing free fatty acids for cells and affecting the maturation of circulating lipoproteins. In this study, the sequence ofLPL gene was obtained by homology-based cloning strategy in black Muscovy duck, and the mRNA expression level of LPL gene, in the development process of muscle tissue, were analyzed by Real-time PCR. The result indicated that the cloned cDNA of LPL gene from black Muscovy duck were 1515 bp in length which con- mined 1485 bp of CDS, encoding 494 amino acids. The nucleotide and amino acid sequence homology of black Muscovy duck were over 90% to poultry LPL gene, and about 70%-75% homologous to human and mammals. The results of Real-time PCR showed that the mRNA expression level of LPL gene was relatively lower in 2-week-old black Muscovy duck. The expression level gradually increased to the highest value, and then declined and main- mined at a low level. The regulation of LPL mRNA expression in male duck＇s leg muscle was the same as that in breast muscle. The expression ofLPL mRNA fluctuated in female duck＇s leg muscles.