【摘要】 目的:为了进一步深入了解BDNF的生物作用,开展基因工程以及应用基础的研究,需要大量的BDNF。因此构建原核表达重组体就显得非常重要。方法:从本室已构建的pGEMBF克隆中获取人BDNF全长编码片段与原核表达载体pGEX-5T连接,构成p5TBF重组体后,经转化大肠杆菌和IPTG诱导。结果:该重组体经蛋白质含量测定,免疫狭缝杂交和ELISA测定,表明该茵体裂解液具有人BDNF抗原活性。结论:p5TBF重组体在大肠杆菌中表达成功。更多还原

【Abstract】 The human brain derived neurotrophic factor (hBDNF), full coding fragment was isolated from our previous constracted pGEMBF clone and linked up with procuryotic expression vector pGEX-5T, then transformation of E. Coli with above recombinant and induced E. Coli with IFFG. The product of hBDNF protein was estimated by three methods respectively-qutitative determination of the total protein concentration in E. Coli celllysate by CCB-G25 colormetric method, by immuno-slot of BDNF in the same lysate and by ELISA of BDNF protein in the same lysate. The estimation result has shown that the hBDNF expression in E. Coli successfully. At last, discussed some open queations of the clinical use of hBDNF protein.更多还原