Figure 1
From: Effect of the silencing of the Ehcp112 gene on the in vitro virulence of Entamoeba histolytica
Expression and purification of rEhCP112 and antibodies generation. (A) Expression and purification of rEhCP112. An Ehcp112 gene fragment (encoding the pro-peptide and the mature enzyme) cloned in the pTrcHIs C expression vector was expressed in E. coli. Then, the rEhCP112 was purified through Ni2+-affinity columns and analyzed by SDS-PAGE. Lane M, molecular weight markers; lane 1, uninduced bacteria extracts; lane 2, bacteria extracts induced by IPTG; lane 3, purified proteins corresponding to the pro-peptide and mature enzyme (52 and 43 kDa). (B) Antibodies generation. The rEhCP112 was inoculated in rabbits. Then, serum was tested by Western blot assays using the rEhCP112 in a SDS-PAGE. (C) Western blot on E. histolytica extracts. To analyze the specificity of the antibody against rEhCP112 we performed Western blot assays on total extracts of E. histolytica trophozoites; Lane 1, control using the preimmune serum; lane 2, antibody against rEhCP112.