Title :
Unleashed DNA production for transfection, vaccines, or labeling
Author :
Domach, M.M. ; Khan, S.A. ; Ataai, A.A.
Author_Institution :
Dept. Chem. Eng., Carnegie Mellon, Pittsburgh, PA, USA
Abstract :
To understand the limits of DNA synthesis and to improve the amount of plasmid DNA produced per growth process, we deregulated the replication of a plasmid that is not subject to antibiotic selection. To understand further what occurs within a host such as E. coli when deregulated plasmid replication occurs, the proteomes of wild-type and transformed cells were contrasted. We find the copy numbers of 15,000 or higher can be obtained for a 3.7 kB plasmid. Replication fidelity or host growth rate are not compromised. The proteomics suggests that among the adaptations are increased Krebs cycle activity and also factors that enable ribosomal assembly and function.
Keywords :
DNA; RNA; cellular biophysics; molecular biophysics; proteins; proteomics; DNA synthesis; E. coli; Krebs cycle activity; antibiotic selection; growth process; host growth rate; plasmid DNA; plasmid replication; proteomes; proteomics; replication fidelity; ribosomal assembly; transfection; unleashed DNA production; vaccines; wild-type cells; Antibiotics; Assembly; DNA; Proteins; Proteomics; RNA; Vaccines;
Conference_Titel :
Biomedical Engineering Conference (NEBEC), 2015 41st Annual Northeast
Conference_Location :
Troy, NY
Print_ISBN :
978-1-4799-8358-2
DOI :
10.1109/NEBEC.2015.7117038