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| Molecular Cloning, Sequence and Expression Characterization of Myosin Heavy Chain 1 Gene (MyHC1) in Goose (Anser anser) at Embryonic Period |
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Abstract Myosin is the major component of myofibrillar thick wire, and plays important roles in the growth and muscle contraction. In this study, reverse transcription PCR and rapid amplification cDNA ends (RACE) methods were used to obtain the full-length cDNA of myosin heavy chain1 gene (MyHC1) in Anser anser. The bioinformatic analysis and expression patter of MyHC1 mRNA at embryonic period detected by Real-time fluorescent quantitative PCR (qRT-PCR) were performed, respectively. The results showed that the MyHC1(GenBank No. KM675469) cDNA full-length was of 6 028 bp including a 5 823 bp open reading frame, 57 bp 5'UTR and 148 bp 3'UTR, and encoded a polypeptide of 1 940 amino acids. The MyHC1 protein was predicted to compose of 31 478 atoms with the formula of C9746H15817N2753O3096S66 and a kind of unstable and hydrophilic protein, with a calculated relative molecular mass of 223 kD, pI of 5.62 and an average hydropathicity of -0.786. The 3-dimensional structure of MyHC1 protein prediction online between Anser anser and Gallus gallus was highly similar, many spiral and folded pieces gathered into globular head with a long fibrous alpha helix tail. Multiple sequence comparisons showed that Anser anser MyHC1 had high homology with Gallus gallus at 92.86%, about 84% with most mammals, and relatively low with amphibians. Further amino acid sequence comparisons showed high homology with Gallus gallus at 96.45%, about 90% with most mammals, and relatively low with Xenopus laevis at 78.13%. Then phylogenetic relationship analysis indicated that mammals formed a large branch, and amphibians formed another branch. Anser anser clustered with Gallus gallus which was similar to the results of the biological classification, which verified they belonged to related species. qRT-PCR result showed that MyHC1 expressed form the early 7 d, increased until 15 d, reached a peak later, then decreased and gradually stabilized after 25 d, showing MyHC1 mRNA expression level of a downward trend after the first rise. This study provides the full-length cDNA sequence and characteristics of geese MyHC1 gene and protein, suggesting that it plays an important role in the process of muscle growth which sipplies the basic information for its function and goose meat quality traits research.
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Received: 08 October 2014
Published: 02 February 2015
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DU Fei, YI Zhi-Xin, YU Hai-Ying, ZHANG Yan, LI Hui-Min, YANG Li, WANG Yu-Zeng, JIANG Dong-Mei, KANG Bo. Effect of Spermine on Expression of Key Genes Related to Polyamine Metabolism and Polyamine Contents in the Duodenum of Female Geese (Anser cygnoides)[J]. 农业生物技术学报, 2019, 27(8): 1445-1451. |
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