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试验旨在研究结核分枝杆菌Rv2626c蛋白对小鼠巨噬细胞RAW264.7细胞凋亡的影响。根据GenBank数据库中结核分枝杆菌Rv2626c基因序列设计引物,并以结核分枝杆菌国际标准株H37Rv cDNA为模板,PCR扩增Rv2626c基因并克隆至慢病毒表达载体pLEX-EGFP中,包装慢病毒并感染RAW264.7细胞,使用Western blotting和流式细胞仪技术检测Rv2626c蛋白表达水平和RAW264.7细胞凋亡率变化。结果显示,成功构建慢病毒表达载体pLEX-EGFP-Rv2626c;成功包装慢病毒并感染RAW264.7细胞;Rv2626c蛋白在RAW264.7细胞中高水平表达显著促进了细胞凋亡。本试验结果表明,在RAW264.7细胞中过表达结核分枝杆菌Rv2626c蛋白能显著性增加其凋亡水平。
The aim of this experiment was to investigate the effect of Mycobacterium tuberculosis Rv2626c protein on the apoptosis of murine macrophage RAW264.7 cells. The primers were designed according to the sequence of Mycobacterium tuberculosis Rv2626c in GenBank and the Mycobacterium tuberculosis H37Rv cDNA was used as a template to amplify the Rv2626c gene and cloned into the lentiviral expression vector pLEX-EGFP. The lentivirus was packaged and infected RAW264.7 cells were used to detect the expression of Rv2626c protein and the apoptosis rate of RAW264.7 cells by Western blotting and flow cytometry. The results showed that the lentivirus expression vector pLEX-EGFP-Rv2626c was successfully constructed, the lentivirus was successfully infected and infected with RAW264.7 cells, and the high expression of Rv2626c protein in RAW264.7 cells significantly promoted apoptosis. The results showed that overexpression of Mycobacterium tuberculosis Rv2626c protein in RAW264.7 cells significantly increased the level of apoptosis.