Rhodobacter capsulatus Gene Transfer Agent (RcGTA) Activity Bioassays

Materials and Reagents

1. GTA donor strain (see Notes 1)
   
2. Recipient strain (see Notes 1)
   
3. RCV broth (Beatty and Gest, 1981)
   
4. YPS broth (Wall et al., 1975)
   
5. YPS agar (2 plates per recipient-donor combination, one plate per donor, and one plate per recipient)
   
6. 0.22 μm filtered GTA buffer (Solioz and Marrs, 1977)
   
7. 0.45 μm low protein binding (PVDF) syringe filters (e.g. EMD Millipore, catalog number: SLHV033RB )
   
8. Plate spreader (Including 95% ethanol + flame)
   
9. 1 ml syringes
   
10. Test tubes for aerobic culturing (e.g. Fisher brand brand, catalog number: 14-961-30 ) w/caps
    
11. Culture tubes for anaerobic/photosynthetic culturing (e.g. Fisher brand, catalog number: 14-959-37A w/screw caps)
    
12. Polypropylene sterile culture tubes (e.g. Simport, catalog number: T405-2A )
    
13. Optional (if transferring photosynthesis marker): Chambers/packs for anaerobic plate growth
14. RCV Medium (see Recipes)
15. YPS Medium (see Recipes)
16. GTA buffer (see Recipes)
    

Equipment

1. Shaking incubator
   
2. Microcentrifuge
   
3. Incandescent light-box or light incubator
4. 1 ml pipettes
5. 1.5 ml Microcentrifuge tubes
   

Procedure