Recombinant Coxiella burnetii Chaperonin GroEL (groEL), partial

The process of generating recombinant Coxiella burnetii 60 kDa chaperonin (groL) in E. coli begins with co-inserting the target gene into an expression vector with an N-terminal 10xHis-tag and C-terminal Myc-tag gene, which is introduced into E. coli cells. The target gene encodes the partial Coxiella burnetii groL protein (326-502aa). These cells are cultured and induced for protein expression. After growing, the cells are lysed to extract the recombinant protein. The collected protein is purified by affinity chromatography. The purity of the protein is evaluated using SDS-PAGE, exceeding 85%.

Coxiella burnetii 60 kDa chaperonin (GroEL) is a type I chaperonin that plays a crucial role in protein folding processes [1]. The 60 kDa chaperonins, including GroEL, are double-ring structures composed of multiple 60-kDa subunits and are involved in folding nascent, translocating, and stress-denatured proteins [2]. These chaperonins are essential for aiding the folding of denatured or newly synthesized polypeptides [3].

References:
[1] C. Wagner, I. Lu, M. Hoffman, W. Sun, J. Trent, & J. Connor, T-complex polypeptide-1 interacts with the erythrocyte cytoskeleton in response to elevated temperatures, Journal of Biological Chemistry, vol. 279, no. 16, p. 16223-16228, 2004. https://doi.org/10.1074/jbc.m310730200
[2] G. Levy‐Rimler, P. Viitanen, C. Weiss, R. Sharkia, A. Greenberg, A. Nivet al., The effect of nucleotides and mitochondrial chaperonin 10 on the structure and chaperone activity of mitochondrial chaperonin 60, European Journal of Biochemistry, vol. 268, no. 12, p. 3465-3472, 2001. https://doi.org/10.1046/j.1432-1327.2001.02243.x
[3] D. Ruggero, A. Ciammaruconi, & P. Londei, The chaperonin of the archaeon sulfolobus solfataricus is an rna-binding protein that participates in ribosomal rna processing, The Embo Journal, vol. 17, no. 12, p. 3471-3477, 1998. https://doi.org/10.1093/emboj/17.12.3471