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| cDNA Cloning, Sequence Structural Analysis and Tissue Expression of Tail-interacting Protein 47 Gene (TIP47) in Xinong Saanen Dairy Goat |
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Abstract Tail-interacting protein 47 gene(TIP47) is important for lipid droplet biogenesis and metabolism, to study the function and expression of goat TIP47 gene, we cloned TIP47 from Xinong Saanen goat(Capra hircus) mammary gland total RNA using RT-PCR and RACE PCR. The results showed that the cDNA sequence of goat TIP47 was 1 906 bp in length (GenBank accession No. HQ846827), including 82 bp of 5'UTR, 1 314 bp of CDS, and 510 bp of 3'UTR, encoding a protein of 437 amino acids with a molecular weight of 47.36 kD and pI 5.74. Nucleotides and amino acid(aa) sequences alignment showed that the nucleotides and aa of goat TIP47 had high score similarity with bovine (Bos taurus), human (Homo sapiens), mouse (Mus musculus) and pig (Sus scrofa) in GenBank. Primary protein structure analysis showed that TIP47 did not contain signal peptide sequence and transmembrane structure; the N-terminal of the protein was more hydrophobic than that of the C-terminal; the C-terminal of the protein was shaped like an L in tertiary structure. Goat TIP47 had the closest relationship with bovine, followed in pig and human, which was generated by phylogenetic tree analysis. TIP47 was expressed in all 12 tissues examined by Real-time fluorescence quantitative PCR, the result showed that the TIP47 mRNA expression in mammary gland was the highest and far higher than those of all other tissues examined, followed in spleen and liver; TIP47 expression in muscle was the lowest. The results suggest that TIP47 plays a potentially role in biogenesis and metabolism of lipid droplet in goat mammary gland, setting the base for function studies of this gene in lipid droplet activites.
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Received: 10 March 2011
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