CPP32在细胞编程死亡调控中起重要的作用.利用CPP32专一性引物,用RT-PCR方法从人鼻咽癌CNE细胞中扩增出约830 bp的片段,经序列分析证明与已发表的CPP32序列完全一致.将扩增出的编码人全长CPP32的cDNA片段克隆入pGEX-2T中,转化大肠杆菌DH5α.转化菌经诱导表达出较高含量的GST-CPP32融合蛋白.进一步研究显示,细菌中表达的CPP32蛋白能自我切割,而且能裂解体外翻译的PARP,从而证明其具有生物活性.
CPP32 has been recently reported to be involved in the early process of programmed cell death. To further study CPP32 and its regulation in the cell, a 830 bp cDNA was cloned by RT-PCR from CNE cells encoding the full length human CPP32 protein and high level expression was achieved in E.coli by using GST expression system. The results showed that the bacterially expressed CPP32 protein is auto-cleaved and capable of cleaving in vitro-translated PARP, thus is fully functional.
陈亚兵,俞春东,郭本昌,丁梅,郭淑贞,曾定,温龙平. CPP32 cDNA的克隆及其表达和活性检测[J].生物化学与生物物理进展,1998,25(2):151-154
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