Cloning And Spatio-temporal Expression Of MEF2A,MEF2B, MEF2C And MEF2D Genes In Different Tissues Of Xingyi Duck

Xingyi ducks aged from 0 to 150 days were chosen to be the research material.We cloned the complete coding region sequence of the MEF2 A, MEF2 B, MEF2 C and MEF2 D genes which were all members of Myocyte-specific enhancer binding factor2 gene family by RT-PCR combined with A/T clone method. We also made the related bioinformatics analysis. At the same time through the real-time fluorescent quantitative PCR(QRT-PCR) we applied the four gene, and detected the mRNA expression level of 10 tissues which was heart, liver, lung, kidney, duodenum, brain,pectoral muscle, leg muscle, gizzard and glandular stomach.The main research results were as follows:(1)In this study we successfully cloned the MEF2 A, MEF2 B, MEF2 C and MEF2 D genes coding sequence in Xingyi duck. The length of the open reading frame sequence was 1479 bp, 1221 bp, 1104bp(1308bp) and 1548 bp respectively. and they encodes 492, 406, 367(435)and 515 amino acids respectively,and two kinds of shearing mode may be existed in MEF2 C gene.(2) In this research, the CDS sequence of the MEF2 B gene was first obtained, the sequence contained the initial password(ATG) and terminate the password(TAG),and through the sequence alignment we found the sequence had higher homology with other Species. Through the NCBI online prediction the mRNA conservative domain structure was analysed, results showed that it contains MADS-box conserved domains and no HJURP_C.(3) The MEF2 gene family's of the Xingyi duck by real-time detection, we found that all the RNAs of this gene family expressd in 0, 30, 60, 90, 120 and 150 days and they also expressd in a different degree in 10 organizations.According to the statistical analysis, there were significant differences in the relative expression of differentgenes in different tissues, different stages and different genders.Every gene expression had reached significant level in sex, but in different tissue,the male and female duck had different gene expression trend at the same growth phase, and the same tissue also had different gene expression trend at different growth phase. This maybe related with law of the growth and development. At the same growth stage the expression of every gene had significant difference in different tissue.And in the same tissue the expression of every gene had significant difference at different growth phase. The expression trend of different gene were not consistent.This showed that the four gene's expression were not synchronous, which maybe concerned with its different function and the order of expression.