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| 结核杆菌HSP65与IL—12多价DNA疫苗的构建 | |
| 引用本文： | 朱振华,王双桂,潘永杰. 结核杆菌HSP65与IL—12多价DNA疫苗的构建[J]. 中国国境卫生检疫杂志, 2009, 0(2): 97-99 |
| 作者姓名： | 朱振华 王双桂 潘永杰 |
| 作者单位： | 象山出入境检验检疫局;象山县中医院检验科; |
| 基金项目： | 象山县科技局资助项目(062313) |
| 摘 要： | [目的]构建结核杆菌H37RV株HSP65基因与IL-12基因的共表达载体pVIVO2-HSP65-IL-12，并研究其在Vero—E6细胞内表达的可行性，为新一代结核多价DNA疫苗寻找理论依据。[方法]采用PCR技术，从培养的结核杆菌H37RV中抽提HSP65基因，克隆到pVIVO2-MCS上的一个多克隆位点，构建pVIVO2-HSP65，将质粒pORF—IL-12上的IL-12基因经双酶切后，亚克隆到pVIVO2-HSP65上，构建成共表达载体pVIVO2-HSP65-IL-12。并经XbaⅠ／AvrⅡ和NcoⅠ／NheⅠ进行双酶切和测序鉴定；用间接免疫荧光法（IFA）检测HSP65和mIL-12基因在Vero—E6细胞中的表达。[结果]双酶切和测序分析证明HSP65基因和mIL-12基因成功克隆到pVIVO2-MCS上，且方向正确，序列无突变；间接免疫荧光试验结果为阳性。[结论]共表达载体pVIVO2-HSP65-IL-12构建成功，且pVIVO2-HSP65-IL-12可在Vero—E6细胞中获得共表达。 |
| 关 键 词： | 杆菌，结核 HSP65 IL-12 共表达 |
Construction of the Multivalent DNA Vaccine of Mycobacterium Tuberculosis HSP65 and Cytokine IL-12 and its Identification |
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| Zhu Zhenhua,Wang Shuanggui,Pan Yongjie.. Construction of the Multivalent DNA Vaccine of Mycobacterium Tuberculosis HSP65 and Cytokine IL-12 and its Identification[J]. Chinese Journal of Frontier Health and Quarantine, 2009, 0(2): 97-99 | |
| Authors: | Zhu Zhenhua Wang Shuanggui Pan Yongjie. |
| Affiliation: | 1.Xiangshan Entry-Exit Inspection and Quarantine Bureau;Xiangshan315700;China;2.Department of Clinical Laboratory;Xiangshan Traditional Medical Hospital;Xiangshan 315700;China. |
| Abstract: | Objective To construct the recombinant plasmid co-expressioned by heat shock protein 65(HSP65) of mycobacterium tuberculosis and IL-12 to express this plasmid in vero-E6 cell for providing evidence of the development of a new multivalent DNA vaccine for tuberculosis. Method HSP65 gene was amplified by PCR with specific primers from the genome of mycobacterium tuberculosis H37RV and IL-12 gene was digested from plasmid Porf-IL-12 with restriction endonucleases NcoI and NheI . The products were further insert... |
| Keywords: | Tuberculosis mycobacterium DNA vaccine Interleukin 12 Heat shock protein 65 Gene co-expression |
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