【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。更多还原

【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 782 ISSN1000-3061 CN11-1998/Q Chin J Biotech May 25,2011 Vol.27 No.5 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.更多还原