Cloning And Sequence Analysis Of Manchurian Wapiti MSTN Gene And Polymorphism Studies

Myostatin (MSTN) is a kind of newly discovered growth differentiation factor, mainly expressed in animal skeleton muscles, which is taking an important role in the growth and regeneration of animal muscles. The deficiency of this gene would cause the increasing of the quantity animal skeleton muscles fiber and becoming fleshy of muscle cells, there is latent using value on flesh deer. The study about deer MSTN gene was rarely reported in the present reports, the all sequence of MSTN gene, bioinformatics research and polymorphism analysis was never reported before. In this thesis, full sequence of Manchurian Wapiti MSTN gene was cloned and sequenced. The sequence structure, nucleotide composition and amino acid were analyzed by bioinformatics methods. Furthermore, mature region DNA sequences of Manchurian Wapiti MSTN were analyzed by PCR-SSCP. The results showed:The three exons of Manchurian Wapiti MSTN gene was cloned and sequenced and submitted in GenBank (accession EF629535). The result showed: the size of the first exon was 373bp, the size of the second exon was 374bp, the size of the third exon was 381bp. The cDNA was 1128bp, it can be inferred that there was 375 amino acids in Manchurian Wapiti MSTN former protein.Secondly, the analysis of the sequence structure and primary, secondary and third-class structure of MSTN gene showed that:①The analysis of homologies of amino acid showed that the inferred amino acid sequence has the character of TGF-βfamily.②The molecular evolution trees of MSTN gene showed that the distance between pig, ovine, goat, dog were the nearest, between birds was nearer and between fish was the most far.③The analysis of physics and chemical character of MSTN gene indicated that the protein molecular quantity was 42. 8KD, isoelectric point was 6. 02.④The analysis of secondary structure of protein showed that there were 73 a-helix, 100 p-sheet and 202 irregular coiled peptide.⑤Homology model the third-class structure of this sequence with a 2arvb as template which has 42% homology to it, the two sites are close in the space.The result of PCR-SSCP analysis for mature region DNA sequences of Manchurian Wapiti MSTN gene indicated that there were polymorphism in mature region DNA sequences Site. Analyzed by X~2 test, the mutation of mature region DNA sequences of Manchurian Wapiti MSTN gene was in balance condition;They were in low polymorphism in PIC mutation site in Manchurian Wapiti by other analysis.