摘要
将本室构建的重组质粒pGEX-5X-TRAIL55-281转入大肠杆菌JM109和HB101中,鉴定后诱导表达,然后针对培养温度、培养时间及诱导剂IPTG浓度设立梯度试验,发现重组质粒pGEX-5X-TRAIL在JM109与HB101中的表达情况相近,可溶性的人GSTr-hTRAIL55-281最佳表达条件为26℃,0.06mmol/L IPTG,200r/m in.利用亲和层析法纯化出大量可溶性GSTr-hTRAIL55-281后,通过Western Blot及流式细胞仪检测发现GSTr-hTRAIL55-281有较好的免疫学活性与生物学活性.为今后TRAIL作为抗肿瘤药物的开发做必要的准备.
pGEX- 5X- TRAIL55-281 was transformed into E. coli strain HB101 and JM109, the recombinant plasmid pGEX- 5X- TRAIL55-281 was identified, and the expression of it was induced by IPTG. And then Gradient tests of cultivate time, temperature and concentration of revulsant IPTG were performed. The resuits showed： The expression of pGEX -5X - TRAIL recombinant plasmid was similarly in E. coli strain HB101 and JM109. The optimum condition for the expression of soluble GST-rh TRAIL55-281 was 26 ℃, 0.06 mmol/L IPTG, 200 r/min. After GST- rh TRAIL was purified through affinity chromatography, it＇ s immunological activity was tested by Western Blot and it＇s apoptosis- inducing activity was tested by flow cytometry. The soluble GST-rh TRAIL55-281 had good immunological and apoptosis -inducing activity. All of these provide a basis for further antineoplastics research.
出处
《生命科学研究》
CAS
CSCD
2005年第3期267-271,共5页
Life Science Research
