摘要
目的体外克隆、表达视网膜神经节细胞分化调控因子Math5,并通过亲和层析的方法进行纯化获得重组表达的Math5蛋白。方法采用限制性内切酶双酶切系统将Math5基因克隆到表达载体pET28a上,并通过载体自身所带6×His标签,利用Ni-NTA纯化出相应的重组蛋白。结果 Math5基因432bp的序列成功克隆到pET28a载体上,通过Ni-NAT纯化出重组表达的Math5蛋白,其纯度为90%,浓度为1.0mg/mL。结论视网膜神经节细胞分化调控因子Math5蛋白的体外表达与纯化为体外蛋白诱导神经前体细胞向视网膜神经节细胞的分化提供蛋白。
Objective To obtain the recombinant expression of Math5 protein by cloning and expressing retinal ganglion cell differentiation control factor Math5 in vitro with the affinity chromatography method. Methods The restriction endonuelease system was used to clone Math5 gene into the expression vector pET28a, and Ni-NTA was used to purify out corresponding recombinant protein by 6 × His tags carried by itself. Results The 432 bp sequence of Math5 gene was successfully cloned into the vector pET28a. The purity of the purified recombinant Math5 protein by Ni-NTA was about 90 % and the concentration was 1.0 mg/mL. Conclusion The expression in vitro and purification of Math5 protein provide the recombinant protein to induce the retinal progenitor cells to retinal ganglion cells.
出处
《中华实用诊断与治疗杂志》
2012年第12期1160-1161,1164,共3页
Journal of Chinese Practical Diagnosis and Therapy
基金
十堰市科技攻关项目(2010STT05)
湖北医药学院研究生启动金(2009QDJ02)
