Haemonchus contortus: siRNA mediated knockdown of matrix metalloproteinase 12A (MMP-12) results in reduction of infectivity

摘要

Cloning, expression, purification and western blot analysis of Hc-MMP-12. Agarose gel electrophoresis of the PCR products. Lane M: DNA molecular marker; Lane 1: the PCR generated fragment of MMP-12 gene with molecular size of 1275 bp. SDS-PAGE. Lane M: standard molecular weight protein marker; Lane 1: expression before IPTG induction; Lanes 2–6: expression after IPTG induction at different time points; Lane 7: pET32a protein before purification; Lane 8: purified pET32a protein. Purification of recombinant MMP-12. Lane 1: recombinant MMP-12 protein before purification; Lane 2: recombinant MMP-12 protein after purification. Western blot analysis of rHc-MMP-12. Lane 1: serum sample from experimentally infected goat; Lane 2: serum sample from normal goat. Western blot analysis of naïve protein. Lane 1: serum sample from SD rats containing polyclonal antibodies against rHc-MMP-12; Lane 2: serum sample from normal SD rats