一种KBMA炭疽疫苗候选株的研制
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十一五国家重大传染病专项 (No. 2008ZX10004-105) 资助。


Development of a killed but metabolically active anthracis vaccine candidate strain
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National Infectious Diseases Major Project of China during the 11th Five-Year (No. 2008ZX10004-105).

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    摘要:

    炭疽病是由炭疽芽胞杆菌Bacillus anthracis引起的一种人畜共患传染病,严重影响着人类的健康。近年来在细菌疫苗的研究中发现一种特殊的现象:细菌被杀死后,体内的代谢活性却仍然维持 (Killed but metabolically active,KBMA)。此发现为炭疽新型疫苗候选株的研制提供了新思路。先通过同源重组的方法,利用pMAD质粒和Cre-loxP重组酶系统完成对缺失两个毒性大质粒的炭疽芽胞杆菌减毒株AP422的uvrAB基因的敲除,得到AP422△uvrAB菌株,然后通过光化学处理 (包括长波紫外光的照射和8-甲氧基补骨脂素处理),使炭疽芽胞杆菌AP422△uvrAB失去繁殖能力。利用四氮唑化合物MTS检测其代谢活性,表明光化学处理杀死后的炭疽芽胞杆菌AP422△uvrAB在至少4 h内维持一个很高的代谢活性水平,即具备典型的KBMA特性。炭疽杆菌AP422△uvrAB的KBMA菌株的成功研制为我们提供了一种新型炭疽疫苗候选株。

    Abstract:

    Anthrax is a zoonosis caused by Bacillus anthracis, which seriously affects human health. In recent years, a special phenomenon is found that the metabolic active of a bacterium remains after it is killed. To development of a KBMA (killed but metabolically active) Bacillus anthracis vaccine candidate strain, a plasmid pMAD and a recombinase system Cre-loxP were used to knockout the uvrAB gene of B. anthracis AP422 which lacks both of two plasmids pXO1 and pXO2. The results of PCR and RT-PCR shows that uvrAB genes were deleted from B. anthracis AP422 chromosome successfully. The constructed B. anthracis AP422△uvrAB was inactivated by photochemical treatment (PCT) including an exposure in a long-wave-length ultraviolet (UVA) light and a treatment of 8-Methoxypsoralen (8-MOP), then the metabolic activity were detected by the method of MTS. The results showed that the killed B. anthracis AP422△uvrAB maintained a highly metabolic activity for at least 4 hours, showing a state of KBMA. The KBMA strain of B. anthracis AP422△uvrAB provides the prospective vaccine candidate strain for anthrax.

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沈非,袁盛凌,展德文,王艳春,任敏,陶好霞,王芃,王令春,陈冬生,刘纯杰. 一种KBMA炭疽疫苗候选株的研制[J]. 生物工程学报, 2011, 27(5): 781-789

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  • 收稿日期:2010-12-13
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