Growing Culture . Transformed cells were inoculated into 3 ml Lysogeny Broth (LB) medium supplemented with chloramphenicol and shaken at 37C for 24 hours. Growing culture for protein purification were inoculated in 100 ml LB medium instead.
Molecular Cloning The parts were synthesized by IDT and cloning was done with the standard BioBrick cutsites.
Flag-tag Protein Purification We used the FLAG octapeptide and Anti-FLAG M2 magnetic beads to extract protein products from the bacteria. Look here for the full
protocol.
See our Lab Notebook!
References
- Ross P et al. (1991) Cellulose Biosynthesis and Function in Bacteria.Microbiological Reviews 55: 35-58. PMID: 2030672
- Milda E. Embuscado, Jay S. Marks, James N. BeMiller, Bacterial cellulose. I. Factors affecting the production of cellulose by Acetobacter xylinum, Food Hydrocolloids, Volume 8, Issue 5, October 1994, Pages 407-418, ISSN 0268-005X, http://dx.doi.org/10.1016/S0268-005X(09)80084-2. (http://www.sciencedirect.com/science/article/pii/S0268005X09800842)
