TY - JOUR
T1 - Identification of Escherichia coli strains for the heterologous overexpression of soluble Clostridium difficile exosporium proteins
AU - Brito-Silva, Christian
AU - Pizarro-Cerda, Jaime
AU - Gil, Fernando
AU - Paredes-Sabja, Daniel
N1 - Funding Information:
This work was supported by grants from Comisión Nacional de Ciencia y Tecnología de Chile (FONDECYT Grant 1151025 ) to D.P-S. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Funding from Millennium Science Initiative of the Ministry of Economy, Development and Tourism, grant “Nucleus in the Biology of Intestinal Microbiota”.
PY - 2018/11/1
Y1 - 2018/11/1
N2 - Clostridium difficile infections are one of the leading causes of hospital-acquired infections. C. difficile spores are considered the morphotype of transmission and recurrent infection due to its natural spore resistance properties. The outermost spore layer, the exosporium, provides the first contact with the environment and the host. However, molecular biology studies on exosporium proteins are lacking primarily due to difficulties in over-expressing these proteins under soluble conditions. In this work, we have developed a protocol to express soluble exosporium proteins of C. difficile spores in the heterologous Escherichia coli host. We found that the optimum soluble expression conditions may vary between 21, 30 and 37 °C, depending on the protein, and at least CdeC, BclA1 and BclA3, required E. coli strains that provided an oxidative environment such as Shuffle T7. These results will allow further studies with recombinant proteins of the exosporium of C. difficile spores.
AB - Clostridium difficile infections are one of the leading causes of hospital-acquired infections. C. difficile spores are considered the morphotype of transmission and recurrent infection due to its natural spore resistance properties. The outermost spore layer, the exosporium, provides the first contact with the environment and the host. However, molecular biology studies on exosporium proteins are lacking primarily due to difficulties in over-expressing these proteins under soluble conditions. In this work, we have developed a protocol to express soluble exosporium proteins of C. difficile spores in the heterologous Escherichia coli host. We found that the optimum soluble expression conditions may vary between 21, 30 and 37 °C, depending on the protein, and at least CdeC, BclA1 and BclA3, required E. coli strains that provided an oxidative environment such as Shuffle T7. These results will allow further studies with recombinant proteins of the exosporium of C. difficile spores.
KW - BclA
KW - C. difficile
KW - CdeC
KW - Exosporium proteins
UR - http://www.scopus.com/inward/record.url?scp=85054804735&partnerID=8YFLogxK
U2 - 10.1016/j.mimet.2018.10.002
DO - 10.1016/j.mimet.2018.10.002
M3 - Article
AN - SCOPUS:85054804735
SN - 0167-7012
VL - 154
SP - 46
EP - 51
JO - Journal of Microbiological Methods
JF - Journal of Microbiological Methods
ER -