[Objective]The objective of this study was to clone the full-length sequence of the coding sequence (CDS) region of TMEM182 gene in sheep,analyze its structure and the sequence of expression protein using online software,so as to the changes in its expression level in various parts of sheep tissues.[Methods]TMEM182 gene was cloned from sheep tail adipose tissue. The physical and chemical properties,hydrophilicity,transmembrane region,glycoylation site,signal peptide,secondary structure,tertiary structure and protein interaction of TMEM182 protein were predicted by online biological information software. Real-time fluorescence quantitative PCR was used to detect the relative expression levels of TMEM182 gene in 13 kinds of sheep tissue samples,including brain,cerebellum,heart,kidney,spleen,liver,lung,skin,ovary,tail fat,subcutaneous fat,perirenal fat and muscle.[Results]The full length of CDS region of sheep TMEM182 gene was 690 bp,encoding 229 amino acids,and the sequence was highly conserved among different species. Bioinformatics prediction results showed that TMEM182 was a stable hydrophobic protein with transmembrane structure and 3 N-glycoylation sites,no signal peptide. The secondary structure of TMEM182 protein was mainly composed of random coil (38.43％) and 2α-helix (30.57％),while the proportion of extension chain (24.89％) and β-turn (6.11％) was relatively small. Meanwhile,the prediction results of tertiary structure were consistent with those of secondary structure. Real-time fluorescence quantitative PCR results showed that the expression levels of TMEM182 gene in muscle,heart and fat were significantly higher than those in liver,kidney,brain,cerebellum,skin and other tissues(P<0.05).[Conclusion]The CDS region fragment of sheep TMEM182 gene was successfully cloned,and the gene was specifically expressed in sheep muscle and adipose tissues. A theoretical basis was provided for further exploring the molecular mechanism of TMEM182 gene in the process of muscle growth and fat deposition in sheep.
维普
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