The KLO8 gene was PCR amplified and cloned into the prokaryotic expression vector pQE41, expressed as 6× His-tagged fusion protein in M15 E. coli and purified on a Ni-NTA column. (A) Protein expression was checked on a 12% acrylamide gel stained with Comassie blue; lane 1 and 2, bacterial lysates from un-induced or 1 mM IPTG-induced cultures, respectively; lane 3, purified rKLO8 protein; M, Protein ladder. (B) Reactivity of the recombinant protein was confirmed in WB analysis using 10 pooled VL sera or 10 pooled healthy control sera from Sudan, diluted 1∶1000; lanes 1 and 2, lysates from IPTG induced cultures blotted with negative or positive sera, respectively; lane 3, purified rKLO8 blotted with positive sera; M, Protein ladder.