结核分枝杆菌Rv2626c蛋白对RAW264.7细胞凋亡的影响

doi:10.16431/j.cnki.1671-7236.2016.04.006

›› 2016, Vol. 43 ›› Issue (4): 892-898.doi: 10.16431/j.cnki.1671-7236.2016.04.006

结核分枝杆菌Rv2626c蛋白对RAW264.7细胞凋亡的影响

孟露萍¹, 史梦婷², 包海洋¹, 付强³, 史慧君³, 王慧勤², 乔军¹, 张辉², 陈创夫¹

1. 石河子大学动物科技学院, 石河子 832003;
2. 石河子大学生命科学学院, 石河子 832003;
3. 新疆农业大学动物医学学院, 乌鲁木齐 830052

收稿日期:2015-09-09 出版日期:2016-04-20 发布日期:2016-04-27
通讯作者: 张辉, 陈创夫 E-mail:604118228@qq.com;ccf-xb@163.com
作者简介:孟露萍(1990-),女,河北石家庄人,硕士生,研究方向:分子病毒学,E-mail:1106370862@qq.com;史梦婷(1989-),女,新疆石河子人,硕士生,研究方向:动物功能基因组与分子免疫学,E-mail:smt1989tg@gmail.com;包海洋(1989-),男,河北承德人,硕士生,研究方向:结核分枝杆菌蛋白质组学,E-mail:997901424@qq.com
基金资助:
国家科技重大专项项目(2013BAI05B05、2013ZX10003003-002)

Effects of Mycobacterium tuberculosis Rv2626c Protein on RAW264.7 Cell Apoptosis

MENG Lu-ping¹, SHI Meng-ting², BAO Hai-yang¹, FU Qiang³, SHI Hui-jun³, WANG Hui-qin², QIAO Jun¹, ZHANG Hui², CHEN Chuang-fu¹

1. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China;
2. College of Life Science, Shihezi University, Shihezi 832003, China;
3. College of Animal Medicine, Xinjiang Agricultural University, Urumqi 830052, China

Received:2015-09-09 Online:2016-04-20 Published:2016-04-27

摘要/Abstract

摘要： 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。

关键词: 结核分枝杆菌; Rv2626c; RAW264.7细胞; 凋亡

Abstract: The assay was aimed to study the effects of Mycobacterium tuberculosis Rv2626c protein on RAW264.7 cell apoptosis.According to the Rv2626c gene sequence of Mycobacterium tuberculosis in GenBank database, Rv2626c primers were designed.The Rv2626c gene was amplified and cloned into the lentiviral vector pLEX-EGFP.After transfection of lentivirus expressing vector with the helper plasmids into HEK-293T cells, lentivirus were packaged and infected into RAW264.7 cells.Then total protein of the infected cells was extracted.Protein levels of Rv2626c and apoptosis levels of RAW264.7 cells were detected using Western blotting and flow cytometry.The results showed that reconstructed lentivirus expressing vector pLEX-EGFP-Rv2626c and the Rv2626c-overexpressing lentivirus Rv2626c-lv were constructed successfully.The overexpressed Rv2626c protein promoted the RAW264.7 cells apoptosis.Overexpression of Mycobacterium tuberculosis Rv2626c protein significantly promoted apoptosis in RAW264.7 cells.

Key words: Mycobacterium tuberculosis; Rv2626c gene; RAW264.7 cells; apoptosis

中图分类号:

Q786

孟露萍, 史梦婷, 包海洋, 付强, 史慧君, 王慧勤, 乔军, 张辉, 陈创夫. 结核分枝杆菌Rv2626c蛋白对RAW264.7细胞凋亡的影响[J]. , 2016, 43(4): 892-898.

MENG Lu-ping, SHI Meng-ting, BAO Hai-yang, FU Qiang, SHI Hui-jun, WANG Hui-qin, QIAO Jun, ZHANG Hui, CHEN Chuang-fu. Effects of Mycobacterium tuberculosis Rv2626c Protein on RAW264.7 Cell Apoptosis[J]. , 2016, 43(4): 892-898.

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结核分枝杆菌Rv2626c蛋白对RAW264.7细胞凋亡的影响

Effects of Mycobacterium tuberculosis Rv2626c Protein on RAW264.7 Cell Apoptosis

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