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. 2013 Jul 10;89(1):63–67. doi: 10.4269/ajtmh.12-0617

Figure 1.

Figure 1.

(A) Ethidium bromide stain patterns of the Gnathostoma spinigerum complete cDNA encoding a matrix metalloproteinase (MMP)-like protein on a 1.0% agarose gel. An arrow indicates band at approximate of 748 base pairs. Lane 1, DNA size markers (ZipEnzyme, Novosibirsk, Russia) and lane 2, the complete cDNA encoding a MMP-like protein amplified by reverse-transcriptase polymerase chain reaction (PCR) with gene-specific primers. (B) Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the fusion-tagged protein with and without recombinant MMP-like protein expressed in Escherichia coli. Coomassie Brilliant Blue stained (a) and immunoblotting probed with (b) and without (c) an anti- NusA antibody. Lane 1, the fusion-tagged protein containing the recombinant MMP-like protein (102 kDa, arrow) and lane 2, the fusion-tagged protein alone (78 kDa, arrow head). (C) SDS-PAGE analysis of the recombinant MMP-like protein cleaved with recombinant enterokinase (rEK) and the recombinant fused-tagged MMP-like proteins. Coomassie Brilliant Blue stained (a) and immunoblotting probed with sera from pooled gnathostomiasis patients (b) and pooled healthy negative controls (c). Lane 1, the rEK-cleaved MMP-like protein and lane 2, the fusion-tagged MMP-like recombinant protein. A thin arrow indicates expression of the fusion-tagged MMP-like protein (102 kDa). An arrow head indicates 78 kDa band of the fusion-tagged protein alone. A thick arrow indicates band of the rEK-cleaved MMP-like protein (28 kDa).