The invention belongs to the field of animal gene engineering. Seven promoters Six1-P, P1, P2, P3, P4, P5 and P6 with different lengths in the upstream of a specific expression gene of skeletal muscles in pigs are cloned from a pig genome. Nucleotide sequences of the promoters are shown in SEQ ID NO:1; 2; 3; 4; 5; 6; and 7. The lengths of the nucleotide sequences are respectively 1,692, 180, 530, 844, 1,143, 1,135 and 1,494bp. The activity analysis on promoter fragments with different lengths of a pig Six1 gene in three eukaryotic cells shows that the promoters of the pig Six1 gene are promoters which express the muscle specificity; except for the P1, other promoters P2, P3, P4, P5 and P6 all have the capacity of promoting gene expression; and in the promoters P2, P3, P4, P5 and P6, except for the promoter P2, other promoters all show the muscle specificity. The invention discloses the seven promoters, a method for preparing corresponding expression vectors of the seven promoters and identification of the promoter activity of the pig Six1 gene by using a dual-luciferase reporting system.