家蚕凋亡蛋白酶(caspase-1)基因在大肠杆菌中的表达

Expression of Bombyx mori caspase-1 Gene in Escherichia coli

为了研究家蚕凋亡蛋白酶(caspase-1)的原核表达及剪切方式,用PCR法扩增出家蚕caspase-1基因的cD-NA,然后插入pET28a载体,得到阳性质粒pET28a-caspase-1,并转化感受态细胞BL21,用异丙基硫代-β-D-半乳糖苷(IPTG)诱导了caspase-1蛋白的表达。应用SDS-PAGE和Western blot分析对其性质作检测,诱导3 h后caspase-1蛋白得到剪切,5 h开始降解。研究结果表明,caspase-1基因可在大肠杆菌BL21中诱导表达,并且能够进行自我剪切。

To study the prokaryotic expression and modification of silkworm apoptosis protease caspase-1 ,the cDNA of silkworm caspase-1 gene was amplified by PCR and inserted into the plasmid pET28a and a prokaryotic expression vector was obtained to transform BL21 cells, The Caspase-1 was expressed under inducement of IPTG identified by SDS-PAGE and Western blot, The results showed that Caspase-1 product was cleaved after induced with IPTG for 3 hours and began to degrade in 5 hours. It indicated that Caspase-1 product can be expressed and self-cleavaaed in E. coil BL21.