Formaldehyde effects on kanamycin resistance gene of inactivated recombinant Escherichia coli vaccines
- PDF / 267,105 Bytes
- 8 Pages / 547.087 x 737.008 pts Page_size
- 88 Downloads / 192 Views
(0123456789().,-volV) ( 01234567 89().,-volV)
ORIGINAL RESEARCH PAPER
Formaldehyde effects on kanamycin resistance gene of inactivated recombinant Escherichia coli vaccines Rafael A. Donassolo . Marcos Roberto A. Ferreira . Clo´vis Moreira Jr . Lucas M. dos Santos . Emili Griep . Gustavo M. S. G. Moreira . Rafael R. Rodrigues . ˆ ngela N. Moreira . Fabricio R. Conceic¸a˜o A
Received: 10 January 2020 / Accepted: 29 May 2020 Ó Springer Nature B.V. 2020
Abstract Objectives Earlier studies have demonstrated the use of inactivated recombinant E. coli (bacterins), to protect against Clostridium spp. in vaccinated animals. These bacterins have a simpler, safer, and faster production process. However, these bacterins carry expression plasmids, containing antibiotic resistance gene, which could be assimilate accidentally by environmental microorganisms. Considering this, we aimed to impair this plasmids using formaldehyde at different concentrations. Results This compound inactivated the highest density of cells in 24 h. KanR cassette amplification was found to be impaired with 0.8% for 24 h or 0.4% for 72 h. Upon electroporation, E. coli DH5a ultracompetent cells were unable to acquire the plasmids extracted from the bacterins after inactivation procedure. Formaldehyde-treated bacterins were incubated with
R. A. Donassolo (&) M. R. A. Ferreira C. Moreira Jr L. M. dos Santos E. Griep ˆ . N. Moreira F. R. Conceic¸a˜o A Centro de Desenvolvimento Tecnolo´gico, Biotecnologia, Universidade Federal de Pelotas, Rio Grande do Sul, Pelotas, Brasil e-mail: [email protected] G. M. S. G. Moreira R. R. Rodrigues Institut fu¨r Biochemie, Biotechnologie und Bioinformatik, Abteilung Biotechnologie , Spielmannstr, Technische Universita¨t Braunschweig, Braunschweig, Germany
other viable strains of E. coli, leading to no detectable gene transfer. Conclusions We found that this compound is effective as an inactivation agent. Here we demonstrate the biosafety involving antibiotic resistance gene of recombinant E. coli vaccines allowing to industrial production and animal application. Keywords Antibiotic resistance gene Chemical inactivation Recombinant vaccines Selection marker
Introduction Since the advent of recombinant DNA technology in the 1970s, several recombinant protein expression systems have been developed such as mammalian and insect cells, plants, fungi, algae, and bacteria (Clark and Cassidy-Hanley 2005). Among these, the bacterial expression system in Escherichia coli has been the most extensively studied with respect to its cellular mechanisms, allowing for easier genetic manipulations (Huang et al. 2012). In addition, this expression system has a variety of strains and expression vectors that may be adopted to fermentation processes as needed (Huang et al. 2012; Moreira et al. 2014). Hence, E. coli is widely used in industrial processes owing to its fast growth in simple, low-cost media, and
123
Biotechnol Lett
high yield (Vincentelli and Romier 2013; Moreira et al. 2014). Recombinant vac
Data Loading...
