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. 2015 Nov 17;6(1):87–98. doi: 10.1534/g3.115.023069

Figure 2.

Figure 2

CC1G_10077-encoded CCTX2 is nematotoxic. (A) Schematic domain representation of CCTX2 predicting two Ricin-type beta-trefoil domains potentially involved in carbohydrate binding. (B) Expression of soluble CCTX2 in E. coli. The cDNA of CC1G_10077 was cloned, and CCTX2 was expressed in the cytoplasm of E. coli BL21(DE3). Extracts of uninduced (UI) and (I) cells, as well as soluble proteins from induced cells (SP), were run on SDS-PAGE and stained with Coomassie Brilliant Blue. The predicted molecular weight of CCTX2 is 89 kDa. (C) Nematotoxicity of CCTX2. CCTX2-expressing bacteria and control bacteria containing ’empty’ vector (EV), or expressing the previously characterized nematotoxic lectin CGL2, were fed for 48 hr to L1 larvae of Caenorhabditis elegans N2 in order to assess the toxicity of CCTX2 toward nematodes. A Welch’s t-test was computed to test the significance of the differences observed between treatments and the ’empty’ vector control. ***p-value ≤ 0.001. Bars represent the standard deviation calculated for four biological replicates. (D) Phase contrast micrographs of Caenorhabditis elegans N2 fed with E. coli BL21 containing ’empty’ vector (EV), or E. coli expressing CGL2 or CCTX2.