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作者通过大肠杆菌-卡介苗(BCG)穿梭质粒pMV261将鼠白细胞介素2(IL-2)基因引入BCG细胞中,并通过加入BCGa抗原信号序列使表达的IL-2分泌到细胞外.重组BCG与鼠脾细胞共同培养,明显地刺激脾细胞分泌γ干扰素.在试验中,作者构建了两组质粒:一组在质粒pMV261上插入小鼠IL-2基因;另一组在质粒pMV261上插入大鼠IL-2基因.为使表达产物分泌到细胞外及区分BCG产生的重组IL-2与哺乳动物细胞产生的IL-2,在每组重组质粒IL-2基因前加入BCGα抗原信号序列或/和流感病毒血凝素表位标记序列.所有新构建质粒都使用 BCG HSP60启动子进行表达.
The authors introduced the murine interleukin 2 (IL-2) gene into BCG cells via E. coli-BCG shuttle plasmid pMV261 and secreted the expressed IL-2 extracellularly by addition of the BCG A antigen signal sequence. Recombinant BCG and Mouse splenocytes co-cultured and obviously stimulated the spleen cells to secrete gamma interferon.In the experiment, the authors constructed two groups of plasmids: one group inserted the mouse IL-2 gene into the plasmid pMV261; the other group inserted into the plasmid pMV261 In order to make the expression product secreted extracellularly and differentiate the recombinant IL-2 produced by BCG and IL-2 produced by mammalian cells, BCGα antigen signal sequence is added before each recombinant plasmid IL-2 gene or / And the influenza virus hemagglutinin epitope tag sequence.All newly constructed plasmids were expressed using the BCG HSP60 promoter.