CBLB502蛋白原核表达及辐射防护作用验证

Expression of CBLB502 in prokaryocytes and verification of radiation protective effect

目的原核表达并纯化TLR5受体的激动剂CBLB502蛋白，验证其辐射防护作用。方法采用pET28b原核表达系统表达CBLB502蛋白，利用Ni-NTA亲和层析柱亲和层析纯化CBLB502蛋白，经报告基因检测方法结合动物实验评价纯化蛋白活性。结果成功表达并纯化CBLB502蛋白，纯度可达95％以上，能够激活NF—κB报告基因，可以提高9Gy照射小鼠存活率，对照组小鼠全部死亡，CBLB502蛋白预防组全部存活。结论成功表达并纯化CBLB502蛋白，并验证其对小鼠的辐射防护作用。

Objective To obtain the purified CBLBS02 protein as an agonist of Toll like receptor 5 in prokaryocytes and to validate its radiation protective effect. Methods pET28b expression system was used to express CBLBS02 protein, and the Ni-NTA agarose and purification column was used to purify CBLB502 protein. The radiation proective effect of CBLB502 was evaluated by NF-κB reporter gene assay and mice experiments. Results CBLB502 protein was expressed and purified, which could activate NF- κB reporter gene in vitro and improved the livability of irradiated mice by 9 Gy T-ray. Conclusions CBLB502 protein could be successfully expressed and purified using prokaryotic expression system. It has the excellent radiation protective effect in mice.