白介素24融合蛋白的表达、纯化和促伤口愈合功能的鉴定  

Expression,purification and identification of wound healing function of Interleukin-24 fusion protein

作　　者：于偌萱 周虹[1] 梁朋[1,2] 

机构地区：[1]四川大学生命科学学院,成都610064 [2]生物治疗国家重点实验室,成都610064

出　　处：《四川大学学报（自然科学版）》2017年第2期436-440,共5页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然科学基金(81171955)

摘　　要：本研究通过优化小鼠白介素24(mIL-24)基因序列,构建以Fc-tag为标签的mIL-24-Fc融合蛋白.对CHO进行MTX分级加压筛选,获得了稳定、高表达mIL-24-Fc蛋白的阳性克隆,同时优化了CHO细胞无血清驯化与稳定表达蛋白体系.经protein A亲和层析纯化mIL-24-Fc蛋白并利用AP binding实验检测生物学活性后,小鼠皮内注射mIL-24-Fc蛋白和伤口愈合实验证明:注射mIL-24-Fc蛋白后,小鼠表皮产生2~3层细胞增生,伤口愈合速度加快.In this study, the mIL-24-Fc fusion protein tagged with Fc-tag was constructed by optimizing the sequence of murine interleukin 24 （mIL-24） gene. The stable and high expression of mIL-24-Fc protein was screened at different concentrations of MTX, and the serum-free cell culture technique and stable expression system of CHO cells were optimized. MIL-24-Fc fusion protein was purified by protein A affinity chromatography and confirmed by AP binding assay. The result showed that after intradermal injections of mouse, a large number of keratinocytes showed two to three proliferation and in wound-healing process, epidermal hyperplasia was promoted and wound healing process was accelerated by mIL-24-Fc.

关 键 词：白介素24 proteinA 银屑病 伤口愈合 

分 类 号：Q511[生物学—生物化学]