二花脸猪ABCG8基因的蛋白功能与组织表达分析

    Protein Function and Tissue Expression Analysis of ABCG8 Gene in Erhualian Pig

    • 摘要: ABC转运蛋白8(ABCG8)基因与家畜抗逆性相关,为探究猪ABCG8编码的蛋白功能及其在各组织中的表达,通过PCR技术克隆二花脸猪ABCG8基因,使用在线生物信息学方法分析ABCG8质结构与功能,利用qRT-PCR检测其组织差异性表达分析。结果表明:该蛋白子式为C3638H5712N982O1027S35,编码730个氨基酸,分子量为80 761.43 kD,pI值为8.73,GRAVY值为0.071,属于疏水蛋白。该蛋白存在109个潜在的N-磷酸化位点和6个O-糖基化位点,不存在信号肽序列,二级结构主要由α螺旋(43.01%)、β-转角(6.16%)、延伸链(14.52%)和无规则卷曲为(36.30%)组成,主要定位在细胞膜和内质网,是定位于细胞膜上的多次跨膜蛋白,可能与ABCG5、NPC1L1、CYP7A1、NR1H3、SCARB1、SREBF2、APOA1、NR1H2、SQLE、NR1H4等蛋白存在相互作用关系。此外,ABCG8基因在二花脸猪各组织中的表达谱分析表明,ABCG8基因在肝脏组织中的表达量最高,小肠次之,在其他组织中几乎不表达。研究结果为进一步研究ABCG8在猪生长性能和肉质性能调控中的机理研究奠定了基础。

       

      Abstract: ABC transporter protein 8 (ABCG8) gene is related to the resistance of livestock. In order to explore the function of the protein encoded by PIG ABCG8 and its expression in various tissues, the ABCG8 gene of Erhualian pig was cloned by PCR technology, and the structure and function of ABCG8 protein were analyzed by using the online bioinformatics method. Meanwhile, the qRT-PCR technology was used to detect the differential expression of its tissue. The results showed that the molecular formula of this protein was C3638H5712N982O1027S35, encoding 730 amino acids, with the molecular weight of 80 761.43 kD, pI value of 8.73, and GRAVY value of 0.071, which belonged to the hydrophobic protein. There were 109 potential N-phosphorylation sites and 6 O-glycosylation sites, and no signal peptide sequence in the protein. The secondary structure of the protein was mainly composed of α helix (43.01%), β-rotation (6.16%), extended chain (14.52%) and random coil (36.30%), which was mainly located in the cell membrane and endoplasmic reticulum. It was a multi-transmembrane protein located on the cell membrane, and may interacted with the proteins such as ABCG5, NPC1L1, CYP7A1, NR1H3, SCARB1, SREBF2, APOA1, NR1H2, SQLE, and NR1H4, etc. In addition, the expression profile analysis of ABCG8 gene in various tissues of Erhualian pig showed that the expression level of ABCG8 was the highest in the liver tissue, followed by the small intestine, but almost no expression in other tissues. The results laid the foundation for the further research on the mechanism of ABCG8 gene in the performance control of pig growth and meat quality.

       

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