Structural and Functional Characterization of Recombinant Interleukin-10 from Indian Major Carp Labeo rohita

Figure 1

Analysis of expression of recombinant IL-10 of L. rohita. (a) E. coli BL21 (DE3) cells harboring pETLrIL-10 were induced with IPTG. Lanes “U” and “I” represent total cell lysates of uninduced and induced cells, respectively. Lanes S and P represent the soluble and pellet fractions, respectively, of the induced cell lysate prepared in 10 mM Tris-HCl, pH 7.0. Lanes and represent the soluble and insoluble fractions, respectively, of the induced cells lysed in 10 mM Tris-HCl, pH 7.0, containing 0.5 M arginine. (b) Western blot analysis of induced cell lysate prepared in the presence of 0.5 M arginine (lane 1) using anti-His antibody. (c) SDS-PAGE (12%) analysis of purified rLrIL-10 (lane 1). Arrow in Panels (a)–(c) points to rLrIL-10. (d) Analysis of oligomeric state of rLrIL-10. SDS-PAGE analysis of the unboiled (lane 1) and boiled rLrIL-10 (lane 2), prepared in nonreducing and reducing conditions, respectively. The arrow head and arrow point to the dimeric and monomeric forms of the rLrIL-10 in unboiled and boiled samples, respectively. M in all the panels indicates protein molecular weight markers.

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