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.
Original language | English |
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Pages (from-to) | 781-789 |
Number of pages | 9 |
Journal | Shengwu Gongcheng Xuebao/Chinese Journal of Biotechnology |
Volume | 27 |
Issue number | 5 |
State | Published - May 2011 |
Externally published | Yes |
Keywords
- 8-MOP
- Cre-LoxP system
- Gene knockout
- Homologous recombination
- Long-wave-length ultraviolet (UVA)
- MTS
- Metabolically active
- UvrAB genes