Expression, Purification, and Functional Characterization of Atypical Xenocin, Its Immunity Protein, and Their Domains from Xenorhabdus nematophila
Figure 2
SDS-PAGE showing purification of recombinant proteins or protein complexes by Ni-NTA chromatography under native conditions. (a) Purification of xenocin-immunity domain (10 kDa) protein complex. Lane M: protein marker; lanes 1 and 2: purified fractions. (b) Purification of catalytic-immunity domain protein complex. Lane M: protein marker; lanes 1 and 2: purified fractions. (c) Purification of full length immunity protein (42 kDa). Lane M: protein marker; lanes 1 and 2: purified fractions. (d) Purification of hemolysin domain (32 kDa). Lane M: protein marker; lanes 1 and 2: purified fractions. (e) SDS-PAGE showing expression and purification of translocation domains of xcinA gene by Ni-NTA chromatography. Lane M: protein marker; lane 1: induced cells; lane 2: uninduced supernatant; lane 3: induced supernatant; lane 4: Wash 1; lanes 5 to 9: fraction number 2 to 6. Arrow indicates the purified translocation domain. (f) Western blot analysis of translocation domain with purified fraction using anti-His antibodies. Lane M, Prestained protein marker; Lane 1: purified sample. (g) SDS-PAGE showing the purification of translocation-receptor domains of xcinA gene by Ni-NTA chromatography. Lane M: protein marker; lanes 1 to 5: fraction number 3 to 7. Arrow indicates the expression and purification of translocation-receptor domain. (h) Western blot analysis of translocation-receptor domain with anti-His antibodies. Lane 1: induced cells; lane 2: Ni-NTA purified protein; Lane 3: prestained protein marker.
