Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi
Figure 1
Heterologous expression of E. cuniculi C1 Hsp70 protein as a histidine fusion protein in E. coli and Western blot analysis using purified C1 antibodies. A Coomassie stained SDS-PAGE gel ((a), right panel) of uninduced and IPTG induced expression of the recombinant protein shows a 76 kDa protein in the induced lane. A Western blot of the SDS-PAGE gel was performed using histidine-tag-specific antibodies ((a), left panel) confirmed the recombinant protein induction. (b) A single 76 kDa band was detected on 1D Western blot of E. cuniculi and E. intestinalis total spore protein using purified C1 Hsp70 protein antibodies. (c) The C1 (ECU02_0100) and B1 (ECU03_0520) Hsp70-related proteins were identified from a Coomassie stained 2D SDS-PAGE gel of E. cuniculi total spore protein by MALDI-MS analysis of trypsin digested gel spots. (d) Western analysis of the 2D gel using the C1 specific antibodies detected only the C1 protein (d).
