耻垢分枝杆菌表型探究举例.ppt

YBR Green I是一种结合于所有dsDNA双螺旋小沟区域的具有绿色激发波长的染料 * * 耻垢分枝杆菌表型研究方法举例 2014.9.12 OUTLINE ?1. Long-term?survival ?2. Sliding?motility ?3. Biofilm?formation 4. Cell aggregation 5. Colonial morphology 6. Staining by Congo red 7. Staining by Crystal violet 8. Sensitivity to SDS 9. Sensitivity to H2O2 (acidic pH, freezing and thawing, heat) 10. Sensitivity to UV exposure 11. Chromosome segregation and elongation 12. Live/dead staining of strains 13. Interactions between Mycobacterium smegmatis and macrophages Outline 长期存活 Bharati et al., A?full-length?bifunctional?protein?involved?in?c-di-GMP?turnover?is?required?for?long-term?survival?under?nutrient starvation?in?Mycobacterium?smegmatis. Microbiology.?2012,7(158):1415-27.? Background: MSDGC-1, having GAF, GGDEF and EAL domains arranged in tandem, and exhibits both c-di-GMP synthesis and degradation activities, deletion of which severely affects long-term survival under conditions of nutrient starvation. Method: The strains were grown in 7H9 containing 0.02% (w/v) glucose and 0.05% (v/v) Tween 80. The antibiotics were omitted from the culture to rule out any effect that they might have on long-term survival. Bacterial cultures were declumped by vortexing the cells with 0.5 mm glass beads before plating on a 7H9 agar plate. The number of c.f.u. was determined at regular time intervals up to 20 days. 耻垢分枝杆菌表型研究方法举例 滑动性 Method I: Cells from colonies grown on LB plates were inoculated via sterile toothpicks onto motility plates. The plates were incubated at 37 ℃ under humid conditions for 2 to 3 days until a transparent halo surrounding the inoculation point was observed for the wild-type strain. motility plates: M63 salts plates supplemented with 0.2% glucose containing 0.3% agarose Background: There is a perfect correlation between the presence of GPLs on the mycobacterial cell wall of M. smegmatis and colony morphology, sliding motility, and biofilm formation. Recht et al., Glycopeptidolipid?acetylation?affects?sliding?motili