Maintaining Wolbachia in cell-free medium.
In this video protocol, procedures are demonstrated to (1) purify Wolbachia symbionts out of cultured mosquito cells, (2) use a fluorescent assay to ascertain the viability of the purified Wolbachia and (3) maintain the now extracellular Wolbachia in cell-free medium. Purified Wolbachia remain alive in the extracellular phase but do not replicate until re-inoculated into eukaryotic cells. Extracellular Wolbachia purified in this manner will remain viable for at least a week at room temperature, and possibly longer. Purified Wolbachia are suitable for micro-injection, DNA extraction and other applications.
Gamston
Courtney
C
Malaria Research Institute, Bloomberg School of Public Health, John Hopkins University, USA.
Rasgon
Jason
J
D000818:Animals; D001431:Bacteriological Techniques; D002455:Cell Division; D002474:Cell-Free System; D002478:Cells, Cultured; D009033:Culicidae / Q000166:cytology / Q000382:microbiology; D003470:Culture Media; D050296:Microbial Viability; D013559:Symbiosis; D020577:Wolbachia / Q000166:cytology / Q000302:isolation & purification* / Q000502:physiology*
D003470:Culture Media