Cloning, Expression, and Polymerization Assay of FtsZ Protein from Staphylococcus aureus

• Published in: Korean Journal of Microbiology and Biotechnology Vol. 40; no. 3
• Main Authors: Son, S.H., Kookmin University, Seoul, Republic of Korea, Lee, D.Y., Seoul Science High School for Gifted Students, Seoul, Republic of Korea, Kim, Y.J., Seoul Science High School for Gifted Students, Seoul, Republic of Korea, Ko, S.H., Seoul Science High School for Gifted Students, Seoul, Republic of Korea, Cho, S.J., Seoul Science High School for Gifted Students, Seoul, Republic of Korea, Jung, H.C., Seoul Science High School for Gifted Students, Seoul, Republic of Korea, Lee, H.H., Kookmin University, Seoul, Republic of Korea
• Format: Journal Article
• Language: Korean
• Published: 01.09.2012
• Subjects: FtsZ; GTPase; polymerization assay; STAPHYLOCOCCUS AUREUS
• ISSN: 1598-642X
• DOI: 10.4014/kjmb.1205.05013

Cytokinesis is the final stage of cell division, dividing one mother cell into two daughter cells. For the cutting of a plasma membrane during bacterial cytokinesis, a tubulin homolog FtsZ protein is recruited from the cytoplasm to the division site. FtsZ protein polymerizes in a GTP-dependent manner and its N-terminal domain has a GTPase activity. In this study, we have begun to characterize FtsZ from Staphylococcus aureus (SA). Full-length SA FtsZ was cloned into pRSFDuet-1 vector and the clone was transformed into a BL21 (DE3) star cell. The recombinant SA FtsZ protein was purified using Ni-NTA affinity chromatography and dialysis. Using a spectrofluorometer, we showed that SA FtsZ undergoes a GTP-dependant polymerization in vitro. The polymer of the SA FtsZ protein disappeared after a few minutes, suggesting that the polymer is degraded as the GTP is consumed. This assay system may well be applied for inhibitor screening targeting S. aureus FtsZ.