Abstract:The cDNA sequences of porcine MYL4 gene were cloned, and verified by bioinformatics method. The cDNA fragment was 1105 bp with an entire open reading frame (ORF), encoding 197 amino acids. The analysis of sequence revealed that the porcine MYL4 gene was highly homologous with that of other species such as canis lupus familiaris (98.4%), Homo sapiens (95.9%),chimpanzee (95.4%) and rhesus monkey(95.4%) in amino acid. The protein encoded by porcine MYL4 gene had conserved domain ‘EFH’ and ‘FRQ1’. The expression analysis of MYL4 gene was carried out in the fetus skeletal muscle (at 33, 65 and 90 days post coitus, dpc) by real-time quantitative PCR. The results indicated that MYL4 gene was expressed with wave model in both Tongcheng and Landrace pigs. There was significant difference at 65 and 90 dpc, but no significant difference at 33 dpc between Tongcheng and Landrace.