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| 稳定表达结核分枝杆菌38抗原的EL-4细胞株的建立及鉴定 | |
| 引用本文： | 康林,王庆敏,胡振林,周凤娟,王磊,孙树汉.稳定表达结核分枝杆菌38抗原的EL-4细胞株的建立及鉴定[J].第二军医大学学报,2003,24(11):1253-1255. |
| 作者姓名： | 康林 王庆敏 胡振林 周凤娟 王磊 孙树汉 |
| 作者单位： | 1. 河北医科大学解剖教研室,石家庄,050017 2. 第二军医大学基础医学部医学遗传学教研室,上海,200433;海军医学研究所舰艇卫生研究室,上海,200433 3. 第二军医大学基础医学部医学遗传学教研室,上海,200433 |
| 基金项目： | 国家高新技术发展规划(863)课题(2001AA28111) |
| 摘 要： | 将结核分枝杆菌的38抗原基因片段经PCR方法扩增并插入到pcDNA3真核表达载体中，通过脂质体转染EL-4细胞，经G418筛选，用RT-PCR方法和ELISA方法检测整合和表达情况。结果成功地构建了pcDNA3—38抗原重组质粒，转染细胞中可检测到38抗原的存在，PCR扩增也证实38抗原基因已稳定整合于EL-4细胞的染色体中。本实验为今后在小鼠体内检测结核DNA疫苗激发的细胞毒性T淋巴细胞(CTL)反应奠定了基础。 |
| 关 键 词： | 结核分枝杆菌 38抗原 EL-4细胞株 鉴定 疫苗 T淋巴细胞 |
| 文章编号： | 0258-879X(2003)11-1253-03 |
| 修稿时间： | 2003年3月17日 |
Establishment of EL-4 cell line stably expressing 38 antigens of Mycobacterium tuberculosis |
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| KANG Lin,WANG Qing-Min HU Zhen-Lin,ZHOU Feng-Juan,WANG Lei SUN Shu-Han".Establishment of EL-4 cell line stably expressing 38 antigens of Mycobacterium tuberculosis[J].Academic Journal of Second Military Medical University,2003,24(11):1253-1255. | |
| Authors: | KANG Lin WANG Qing-Min HU Zhen-Lin ZHOU Feng-Juan WANG Lei SUN Shu-Han" |
| Abstract: | The 38 antigen cDNA of Mycobacterium tuberculosis was amplified and inserted into pcDNAS eukaryotic expressing vector. The recombinant plasmid was transfected into EL-4 cell line by liposome and screened by G418. The intergra-tion and expression of 38 antigen cDNA was detected by RT-PCR and ELISA respectively. The results showed that recombinant plasmid pcDNA3-38 was successfully constructed. It further confirmed that 38 antigen cDNA had been stably integrated into the chromosome of EL-4 cell by RT-PCR method,and that 38 antigen could express stably in transfected EL-4 cells. This study provided basis for detecting the cytotoxic T-lymphocyte (CTL) reaction stimulated by DNA vaccine of tuberculosis in rats. |
| Keywords: | Mycobacterium tuberculosis 38 antigen EL-4 cell line |
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