论文部分内容阅读
将含全长FascDNA的质粒利用PCR技术进行特定片段删除 ,获得sFascDNA。通过DNA重组技术 ,将sFascDNA插入到反转录病毒载体 pLXIN ,构建了重组表达载体pLXIN sFas。经酶切鉴定及测序 ,表明成功地构建了重组表达载体 pLXIN sFas。该载体经PA317细胞包装后 ,感染靶细胞COS 7,分别采用ELISA及凋亡抑制实验检测其蛋白表达量为 (2 .2± 0 .7) μg/ml,且具有良好的生物学活性 ,能明显抑制抗 Fas(CH 11)介导的T淋巴瘤细胞株Jurkat的凋亡。这一结果为探讨Fas和FasL途径在白血病发生中的作用奠定了基础
The full-length FascDNA plasmid using PCR technology to delete specific fragments to obtain sFascDNA. The sFasc DNA was inserted into the retroviral vector pLXIN by DNA recombination technology to construct the recombinant expression vector pLXIN sFas. After restriction enzyme digestion and sequencing, the recombinant expression vector pLXIN sFas was successfully constructed. After being packaged in PA317 cells, the vector was infected with COS7, and its protein expression was (2. 2 ± 0. 7) μg / ml by ELISA and apoptosis inhibition test, respectively, and had good biological activity. Significantly inhibited apoptosis of anti-Fas (CH 11) -mediated T lymphoma cell line Jurkat. This result laid the foundation for exploring the role of Fas and FasL pathways in the pathogenesis of leukemia