Materials and Reagents

1. R. capsulatus recipient strain
   
2. RCV broth
   
3. RCV agar
   
4. Sucrose
   
5. L-malic acid (Sigma-Aldrich, catalog number: 240176 )
   
6. Thiamine hydrochloride (Sigma-Aldrich, catalog number: T1270 )
   
7. Escherichia coli cloning strain capable of conjugation (e.g. S17-1)
   
8. Suicide plasmid, such as pZJD29A (Z. Jiang and C. E. Bauer, unpublished strain construction), that contains the sacB gene and an antibiotic resistance gene, but no Rhodobacter capsulatus compatible origin of replication. The suicide plasmid pZJD29A is a derivative of pJP5603 and has a RK6-based origin of replication making it a “universal” suicide plasmid (Penfold and Pemberton, 1992).
   
9. Appropriate antibiotic (resistance specified by the suicide plasmid)
10. RCV broth/agar (Beatty and Gest, 1981) (see Recipes)
11. Trace element solution (in 250 ml dH₂O) (see Recipes)
    

Equipment

1. 30 °C shaker
   
2. 30 °C incubator
   
3. Test tubes
   
4. Petri plates
   

Procedure