Gene Isolation, Sequence Features And Tissue-specific Expression Associated With Growth Traits In Silurus Lanzhouensis

The growth and development of the skeletal muscle of animals is regulated by a series of factors in time and space, and Myo D, Myo G and MSTN are three important myogenic determining factors. In order to explore the genes associated with growth traits of Silurus lanzhouensis, coding sequences(CDS) and the whole genome of Myo D, Myo G and MSTN gene in Silurus lanzhouensis were cloned and analyzed using RT-PCR, TA-cloning and nucleotide sequencing techniques in this study. Expression patterns of these three genes in eight different tissues among different individual sex Silurus lanzhouensis were investigated using real-time fluorescent quantitative method. The main results are as follows:1. The coding sequence(CDS) and complete sequence of Myo D gene(Gen Bank accession number: KT277551 and KT339175, respectively) were obtained with the fragment size of 810 bp and 1210 bp, which contain 5’ UTR(63 bp) and 3’ UTR(58 bp); the open reading frame(ORF) of Myo D gene contains three exons and two introns, the length of three exons are 510 bp, 80 bp and 220 bp, and the length of two introns are 156 bp and 123 bp, respectively. An acid soluble protein with a total of 269 amino acids was encoded by the Myo D gene. Subcellular localization of Myo D was mainly in nucleus(56.5%).The secondary structure of Myo D was a helix loop helix(b HLH). The study is also showed that, Myo D gene can be used to study the evolutionary relationship between S. lanzhouensis and other Siluriformes species. Tissue-specific expression results show that, the expression of Myo D gene in S. lanzhouensis muscle tissue was significantly higher than that of other tissues, and expression of muscle tissue of the male was significantly higher than those of female.2. The complete sequence of CDS with length of 762 bp(Gen Bank accession No.: KT580948) and the whole sequence of Myo G gene with length of 1223 bp(Gen Bank accession No.: KU302770) were obtained by cloning sequence. The ORF contains three exons and two introns, the length of three exons is 539 bp, 104 bp and 119 bp, and the length of two introns is 330 bp and 131 bp. A total of 253 amino acids were encoded by the Myo G gene. Subcellular localization of Myo G was mainly in nucleus(52.2%).The secondary structure of Myo G was a helix loop helix(b HLH). The study is also showed that, Myo G gene can be used to study the evolutionary relationship between S. lanzhouensis and other Siluriformes species. Tissue-specific expression results showed that, the expression of Myo G gene in S. lanzhouensis muscle tissue was significantly higher than that of other tissues and expression of muscle tissue of the male was significantly higher than that of females.3. The complete sequence of CDS with length of 1182 bp(Gen Bank accession No.: KU302769) of MSTN was obtained by cloning sequence, and the ORF includes three exons and two introns. A total of 393 amino acids were encoded by the MSTN gene. Subcellular localization of MSTN was mainly in outside the nucleus(52.2%). The result of secondary structure analysis showed MSTN is a complex type, the predictable third structure is composed of alpha-helix and random coil. The study is also showed that, MSTN gene can be used to study the evolutionary relationship between S. lanzhouensis and other Siluriformes species. In the eight examined tissues, the MSTN gene was highly expressed in muscle, weakly expressed in brain, and there was no obvious expression in other tissues.