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Figure 1: Purification and analysis of TcSP2 and recombinant proteins. A) The TcSP2 gene was amplified by PCR and cloned in frame in the pRSETB plasmid. DNA fragments coding for the CHP and ATPase domains were obtained from the TcHSP70 gene and cloned in frame at the 3 ‘end of TcSP2. The expected sizes of the recombinant proteins rTcSP2, rTcSP2-CHP and rTcSP2-ATP are 30.1, 45.0 and 66.5 kDa, respectively. B) Purified recombinant proteins were analyzed by 12% SDSPAGE and visualized by staining with Coomassie blue. The arrows indicate the location of the recombinant proteins; MW, molecular weight marker. C) Immunoblotting of recombinant proteins. Anti-histidine monoclonal antibodies (1:1000) were used to detect rTcSP2 and anti- TcHSP70 polyclonal antibodies (1:2000) were used to detect rTcSP2-CHP and rTcSP2-ATP. |