Abstract: In present research, the sequences of MyoD gene and 5′-flanking region are amplified from DNA isolated from the largemouth bass by using PCR and GeneWalking technique. A total of 3797 bp sequence of MyoD gene and 5′-flanking region was obtained.The MyoD encoding gene consisted of three extrons（591 bp,81 bp and 78 bp respectively）and two introns（1077 bp and 486 bp）. Sequence analysis of 5′-flanking region of MyoD gene revealed that it contained putative muscle growth response elements such as E-boxes , myocyte enhancer factor 2 (MEF2), muscle-specific Mt binding site (MTBF). TATA boxes, Octamer-binding factor 1(OCT1), activator protein 4 (AP4) and OCAAT box were also detected. 7 SNP sites in introns were identified from MyoD gene by using PCR-SSCP technique and sequencing, however no polymorphism existed in extron and 5′-flanking region. The percentage of SNPs is between 0.042-0.353 in largemouth bass. These SNPs might be useful for a further study on MyoD gene’s relation to production performance in largemouth bass.