摘要
用真核表达引物从pGEM-IL-2重组质粒中扩增出猪IL-2基因,将目的基因和真核表达载体pPIC9K连接转入E.coli的JM109中,得到了猪pPIC9K-IL-2重组表达质粒。通过电激法将经SalⅠ酶切线性化的pPIC9K-IL-2质粒转化到巴斯德毕赤酵母GS115感受态细胞中,利用甲醇诱导表达,经SDS-PAGE电泳分析,表明在摇床水平及发酵罐中均表达出约17 ku大小的分泌性目的蛋白,采用Sephadex G-100分子筛层析对其表达产物进行纯化,纯化结果理想。
The porcine IL-2 gene, which was amplified from pGEM-IL-2 with expression primers, was inserted into the expression vector pPIC9K and then transformed into Escherichia coll. Positive recombinants pPIC9K- IL-2 were selected, sequenced and linearized by Sal Ⅰ. After the linearized DNA of pPIC9K- IL-2 was transformed into Pichia pastoris GS115 by electroporation, the IL-2 gene was expressed by induction of methyl alcohol. SDS-PAGE analysis showed that the IL-2 gene was expressed successfully in Pichia pastoris both in the shake culture and the fermentor, and the porcine IL-2 protein of approximately 17 ku in size was secreted. The expressed protein was successfully by using Sephadex G-100.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2006年第4期315-319,共5页
Chinese Veterinary Science
基金
农业结构调整重大技术研究专项(04-10-03B)
兰州市科技攻关计划项目(05-1-47)
