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 of LPL 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 contained 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 maintained 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 of LPL mRNA fluctuated in female duck’s leg muscles.