摘要
目的构建以绿脓杜菌外毒素A(PE40)为毒性部分的重组毒素,并研究其抗头颈部肿瘤的效应。方法利用基因工程技术将α促黑激素(-αmelanoma stimulating hormone,-αMSH)与绿脓杆菌外毒素A的部分重组基因,克隆到高效表达载体pET-20b中,表达出重组蛋白MSH-PE40,进行头颈部肿瘤细胞的体外杀伤实验。结果成功地将-αMSH基因与PE40基因融合,处于T7启动子和终止子调控之下。初步纯化的MSH-PE40对黑色素瘤具有明显的细胞毒性。结论MSH-PE40具有特异性杀伤-αMSH受体的肿瘤的功能。
Objective To construct anti-tumor recombinant toxin with PE4O as toxic part and study its cytotoxic effect. Method Recombinant gene of α-MSH and PF40 using gene recombinant technique. Coloned the α-MSH-PE40 gene into the expression vec tor pET-20b, induced with IPIG and expressed the recombinant protein. Used the recombinant protein to exam the eytotoxin effect i vitro. Results Analysis of digesting with restriction endonueleases and DNA sequencing verified that the junction between two gene had fight reading frame. Conclusion MSH-PE40 have strong toxin to melanoma cell B16, laryngeal carcinoma cell Hep-2, the control groups have no obvious changes.
出处
《中国实验诊断学》
2005年第5期680-682,共3页
Chinese Journal of Laboratory Diagnosis
