Phage-free production of single-stranded DNA with user-defined sequence using Escherichia coli
Karl Behler, doctoral thesis Technical University of Munich, 2022
Appropriate distribution of the M13 phage genes between helper plasmid and phagemid enabled the scalable production of single-stranded DNA (ssDNA) with user-defined sequence in a fed-batch process with Escherichia coli in a stirred-tank bioreactor without making use of self-replicating phages. This process increased the maximum ssDNA concentration by a factor of 40 compared to literature data, which was also demonstrated on a m3 scale with an overall ssDNA isolation yield of 92%.
Publications
- Behler K, Honemann MN, Santos A-R, Dietz H, Weuster-Botz D (2022): Phage-free production of artificial ssDNA with Escherichia coli. Biotechnol Bioeng 119: 2878-2889.
- Praetorius F, Kick B, Behler KL, Honemann MN, Weuster-Botz D, Dietz H (2017): Biotechnological mass production of DNA origami. Nature 552: 84-87.
- Kick B, Behler KL, Severin TS, Weuster-Botz D (2017): Chemostat studies of bacteriophage M13 infected Escherichia coli JM 109 for continuous ssDNA production. J Biotechnol 258: 92-100.