Liang-shan Semi-fine Wool Sheep Mef2A, Mef2C Gene Clone And Tissues With Time Expression

Mef2 belongs to MADS family member and its binding site sequences generally existence in the control region of specificity express gene in muscle, and has vast influence on muscle and nerve development. Its product has important contribution to cell differentiation,sustain and regeneration for muscle. The MADS region compose by 57 amino acids, and has exceeding conservation.27 amino acids of Mef2 in the end of MADS region, boost affinity of DNA binding,protein dimerization and provid the place of other cofactor interaction.Liang-shan semi-fine wool sheep were used to study the Mef2A and Mef2C gene structure and expression patterns in different tissues at different time after birth. The complete coding region of gene was amplified according to the sequence of conservative parts in different species by GenBank. We use QRT-PCR method to analyses the relative expression levels of Mef2A and MEF2C gene in different tissues at different time by the primer designed on the basis of the cloning sequence. We get parts of Mef2A and complete CDS region of Mef2C sequence. Mef2A contains 779bp and have MADS and MEF2 Kringle, homology with cattle,Mus musculus,human is 98.8%,95.6%,92% respectively. MEF2C gene contains contains 1 302 bp and encodes 434 amino acids. The amino acids sequence of Liang-shan semi-fine wool sheep homology with cattle,Mus musculus,human is 99.5%,93%,99.8% respectively. The resuts tell us Mef2A mutation higher than Mef2C in the evolution process. The N-terminal of the two gene were very conserved after homology blast. The high frequent mutation region behind Mef2 structural domain.The C-terminal of Mef2C was also conserved, and the middle region mutated dramatically. The signal peptide of the two gene were in the same site and inside the MADS region.The cleavage site of the signal peptide was from 29 to 30 amino acids. Ser residus is the major residus in the phosphorylation sites. The secondary structure of protein is dominate by Random coil, and also contains Helix and Sheet. A two-dimentional membrane spaning domain was exist between 90 and 108 amino acids of Mef2A. A two-dimentional membrane spaning domain was exist between 192 and 211 amino acids and two one-dimentional membrane spaning domain from inside to outside. Three hydrophobicity regions of Mef2A contain 9,9,2 amino acids respectively. Nine hydrophobicity regions of Mef2C contain 10,2,12,6,2,2,15,2,11 amino acids respectively.We researched the expression of Mef2A,Mef2C genes in different tisssues with different growth period by QRT-PCR technique. Mef2A is high expression in tissues except skin nevertheless have were significantly difference in different time (P<0.05), so the maximum expression in different time isn't the same tissue.Though the study tell us Mef2A have expression time series during tissues. The Mef2C expression level of cerebra and dorsal muscle was higher than other tissues (P<0.05).The expression patterns of Mef2C were significantly difference in different tissues when the age increased (P<0.05). Through the research we can guess the both of gene expression pattern was tissue-specific and plays an important role in cerebra regulation and growth development after birth.