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将LacZ基因重组于ElA、ElB缺失腺病毒载体中的SRα、EFα或CA启动子下游,应用辅助细胞(293细胞)获得非复制感染性重组病毒,测得病毒滴度为0.42×10~9PFU/ml。包装LacZ基因或GM-CSF基因的逆转录病毒载体产生细胞(CRIP)由美国Richard CM.Mullinan教授惠赠。病毒滴度为0.1-7.7×10~6PFU/ml。应用这两种病毒载体,将外源基因导入人外周血淋巴细胞、人白血病原代细胞与K562、TALL-1细胞以及小鼠白血病细胞M1与WEHI-3B细胞,探索提高基因导入效率和表达效率的试验方法,比较了两种病毒载体基因转移的特点。
The LacZ gene was recombined downstream of the SRα, EFα or CA promoter in ElA and ElB-deficient adenoviral vectors, and non-replicating infectious recombinant viruses were obtained using helper cells (293 cells), and the virus titer was measured to be 0.42×10-9 PFU/mL. Ml. The retroviral vector-producing cells (CRIP) packaging the LacZ gene or the GM-CSF gene were gifted by Prof. Richard CM. Mullinan, USA. The virus titer was 0.1-7.7 x 10-6 PFU/ml. Using these two viral vectors, the introduction of exogenous genes into human peripheral blood lymphocytes, human leukemia primary cells and K562, TALL-1 cells, and mouse leukemia cells M1 and WEHI-3B cells were explored to improve gene transfer efficiency and expression efficiency The test method compares the gene transfer characteristics of two viral vectors.