犬MG53基因的克隆与表达

发布时间：2018-06-25 07:36

本文选题：犬 + MG　；参考：《中国兽医科学》2017年01期

【摘要】：为进一步研究犬MG53在机体内的分布情况及其生物活性,本研究设计特异性引物进行犬MG53目的基因的克隆,并将其连接至p MAL-C5X表达载体中,利用大肠杆菌表达系统进行表达。结果显示,克隆得到由1 435个核苷酸构成的犬MG53基因;获得MG53蛋白与麦芽糖结合蛋白标签(MBP)融合表达的可溶性重组蛋白,大小约为95 ku。通过结构及生物学功能预测发现,该蛋白易降解,具有RING、B-Box、Coiledcoil及SPRY-PRY四个功能结构域,并分布有半胱胺(Cysteamine)、2-氨基乙硫醇(2-Amino-1-Ethanethiol)、硫代乙醇胺(Thioethanolamine)、2,3-Deshydrolanthionine、Zn2+及Mg2+等分子结合位点,具有蛋白结合能力。上述结果表明,犬MG53与人、家兔、小鼠、大鼠MG53的结构相似,并可能具有参与泛素化、骨骼肌细胞修复等重要的生物学功能。
[Abstract]:In order to further study the distribution and biological activity of dog MG53, a specific primer was designed to clone the target gene of dog MG53, which was ligated into pMAL-C5X expression vector and expressed by E. coli expression system. The results showed that the canine MG53 gene composed of 1,435 nucleotides was cloned and the soluble recombinant protein expressed by fusion of MG53 protein and maltose binding protein label (MBP) was obtained, the size of which was about 95 ku. It was found that the protein was easy to degrade and had four functional domains RINGB-Boxen Coiledcoil and SPRY-PRY. The protein was characterized by cysteamine 2-aminoethiol thioethanolamine 2 3-Deshydrolanthionine Zn2 and Mg2. These results suggest that the structure of dog MG53 is similar to that of human, rabbit, mouse and rat MG53, and may have important biological functions such as ubiquitization, skeletal muscle cell repair and so on.
【作者单位】：华南农业大学兽医学院广东省兽医临床重大疾病综合防控重点实验室广东省宠物工程技术研究工程中心;
【基金】：广东省高等学校优秀青年教师培养计划资助项目(YQ2015030) 广东省兽医临床重大疾病综合防控重点实验室资助项目(2013A061401013)
【分类号】：S829.2

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