猪亲环素18的基因克隆与融合蛋白的表达

Molecular Cloning the Pig Cyclophilin 18 (pCyP18) and Expression of the Fusion Protein

亲环素18(CyP18)具有肽脯氨酰顺/反异构酶活性,广泛存在于不同物种中,是一种高度保守的蛋白质.它能和免疫抑制性药物环孢菌素 A 结合,参与免疫抑制;此外,作为分子伴侣,它还加速蛋白质的折叠、组装和分解.从猪肝中提取总 RNA,首次通过 RT-PCR 扩增出 pCyP18,对其基因进行克隆和序列测定,并转入大肠杆菌中进行高效表达,得到纯化的 pCyP18蛋白.

Cyclophilin 18 （CyP18）, a cellular chaperone with cis-trans prolyl isomerase activity is ubiquitous, highly conserved protein that is found in different species in evoluiton. CyP18 binds tightly to the immunosuppressive drug cyclosporin A （CsA） and participates in immunosuppression. Furthermore, CyP18 acts as a molecular chaperone facilitating protein folding and the assembly/disassembly of protein complexes. In this work, we prepared the total RNA from pig liver, amplified pCyP 18 by RT-PCR and clone for the first time this gene in E. coli. After determination of the sequence, the pCyP18 was expressed and the purified protein of pCyP18 was acquired.