农业生物技术学报2011，Vol.19Issue(5)：864-872,9.DOI:10.3969/j.issn.1674-7968.2011.05.012
西农萨能羊47kD尾部相互作用蛋白基因(TIP47)的cDNA克隆、序列结构分析与组织表达
cDNA Cloning, Sequence Structural Analysis and Tissue Expression of Tail-interacting Protein 47 Gene (TIP47) in Xinong Saanen Dairy Goat
摘要
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 (Copra 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 withbovine, 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 T1P47 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.关键词
西农萨能羊/47 kD尾部相互作用蛋白(TIP47)/脂滴形成/基因克隆/序列结构分析/组织表达Key words
Xinong Saanen dairy goat/ Tail-interacting protein 47(TIP47)/ Lipd droplet biogenesis/ Gene cloning/ Sequence structural analysis/ Tissue expression引用本文复制引用
郝娟,罗军,王紫骞,赵旺生,孙雨婷,钟瑜,李君..西农萨能羊47kD尾部相互作用蛋白基因(TIP47)的cDNA克隆、序列结构分析与组织表达[J].农业生物技术学报,2011,19(5):864-872,9.基金项目
本研究由国家转基因生物新品种培育项目(No.2009ZX08009-162B)、陕两省重大科技创新项目(No.2009ZKC07-01-1)和省部级公益性行业(农业)科研专项(No.201103038)共同资助 （No.2009ZX08009-162B）
