| 分泌性表达融合蛋白CFP10-ESAT6重组卡介苗构建研究 |
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| 引用本文： | 张红宇,李晓恒,范兴,罗道泉,吴少庭. 分泌性表达融合蛋白CFP10-ESAT6重组卡介苗构建研究[J]. 中国热带医学, 2009, 9(5): 787-789,840 |
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| 作者姓名： | 张红宇 李晓恒 范兴 罗道泉 吴少庭 |
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| 作者单位： | 1. 深圳市疾病预防控制中心,广东,深圳,518020
2. 深圳市慢性病防治院,广东,深州,518020 |
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| 基金项目： | 深圳市科技计划资助项目(200702112) |
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| 摘 要： | 目的构建分泌表达融合蛋白CFP10-ESAT6的重组卡介苗。方法采用基因拼接（Gene SOEing）法，体外扩增结核杆菌CFP10-ESAT6融合基因，插入大肠杆菌-分枝杆菌穿梭表达质粒pBCG3000，构建重组穿梭表达质粒pBCG3000-CFP10-ESAT6，用电穿孔法将pBCG3000-CFPIO-ESAT6质粒转化BCG细胞，得到重组卡介苗，热诱导表达CFPIO—ESAT6融合蛋白，SDS—PAGE电泳观察CFP10-ESAT6融合蛋白的表达，Western blot鉴定其生物活性。结果经PCR、酶切及测序鉴定，证实成功构建重组质粒pBCG3000-CFP10-ESAT6，经热诱导表达出约22kDa的CFP10-ESAT6蛋白，可分别被鼠抗ESAT6血清、鼠抗CFP10血清识别。结论分泌性表达融合蛋白CFP10-ESAT6的重组卡介苗构建成功。
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| 关 键 词： | 融合蛋白 CFP10 ESAT6 重组卡介苗 穿梭表达质粒 |
Construction, expression and identification of a recombinant BCG vaccine secreting the fusion protein CFP10-ESAT6 |
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| ZHANG Hong -yu,LI Xiao -heng,FAN Xing,et al.. Construction, expression and identification of a recombinant BCG vaccine secreting the fusion protein CFP10-ESAT6[J]. China Tropical Medicine, 2009, 9(5): 787-789,840 |
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| Authors: | ZHANG Hong -yu LI Xiao -heng FAN Xing et al. |
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| Affiliation: | 1 Shenzhen Municipal Center for Disease Control and Prevention;Shenzhen 518020;Guangdong;P.R. China |
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| Abstract: | Objective To construct a recombinant BCG exressing the fusion protein CFP10 -ESAT6. Methods Mycobacterium tuberculosis fusional gene CFP10 -ESAT6 was amplified by Gene SOEing and cloned to an E.coli - Mycobacterium shuttle expressing plasmid pBCG3000, and then the recombinant shuttle expressing plasmid pBCG3000 - CFP10-ESAT6 was successfully constructed. The recombinant plasmid pBCG3000- CFP10-ESAT6 was transduced into BCG by electroporation. Then the recombinant BCG was induced by heat. The expression and ... |
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| Keywords: | Fusion protein ESAT6 CFP10 Recombinant BCG Suttle expression plasmid |
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