论文部分内容阅读
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。
In order to further study the distribution of MG53 in vivo and its biological activity, we designed specific primers to clone MG53 gene and ligated it into pMAL-C5X expression vector and expressed it by using E.coli expression system . The results showed that a canine MG53 gene consisting of 1 435 nucleotides was cloned, and a soluble recombinant protein with a fusion protein of MG53 and maltose binding protein (MBP) was obtained, with a size of about 95 ku. It was found that the protein was easily degraded by the structural and biological function prediction, and had four functional domains of RING, B-Box, Coiledcoil and SPRY-PRY. Cysteamine, 2-aminoethanethiol -Amino-1-Ethanethiol, Thioethanolamine, 2,3-Deshydrolanthionine, Zn2 +, and Mg2 +, which have protein-binding ability. The above results show that the structure of MG53 is similar to that of human, rabbit, mouse and rat MG53, and may have important biological functions such as ubiquitination and repair of skeletal muscle cells.