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目的:获取抗肿瘤坏死因子相关凋亡诱导配体(TRAIL)抗体。方法:利用PCR技术和基因重组技术将TRAIL基因构建于原核表达载体pPreTMH-Tb中,经酶切、测序鉴定证实构建完全正确后,通过IPTG诱导在大肠杆菌DH-5α中获得表达,并将TRAIL蛋白免疫家兔,通过Dot blot及Western blot检测抗TRAIL抗体的产生。结果:成功构建TRAIL基因原核表达载体,并在大肠杆菌DH-5α中获得表达,TRAIL蛋白表达量占菌体蛋白质总量的11.8％,Dot blot及Western blot检测证实获得了抗TRAIL抗体。结论:成功制备抗TRAIL抗体,这为TRAIL在真核细胞水平研究奠定了实验基础。
Objective: To obtain anti-tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) antibody. Methods: The TRAIL gene was constructed by PCR and gene recombination technology. After digestion and sequencing, the TRAIL gene was confirmed to be completely correct and then induced by IPTG in E. coli DH-5α. The TRAIL Protein was used to immunize rabbits and anti-TRAIL antibody production was detected by Dot blot and Western blot. Results: The prokaryotic expression vector of TRAIL gene was successfully constructed and expressed in E. coli DH-5α. The expression of TRAIL protein accounted for 11.8% of the total bacterial proteins. Dot blot and Western blot confirmed the anti-TRAIL antibody. Conclusion: The successful preparation of anti-TRAIL antibody, which TRAIL in eukaryotic cells laid the foundation for the study.