Plasmid construction and transient transfection of U-2 OS cells

A TcoClec ORF (TcIL3000.0.50510) was amplified from T. congolense IL 3000 genomic DNA using the two specific primers TcoClec-Fw (5′-CTCGAGATGAGCAAAGGGAAACACG-3′) and TcoClecSTOP-rev (5′-CTACTGAGCAACCGCCGGCGAC-3′), cloned into pcDNA3.1/CT-TOPO (Invitrogen) and sequenced. Exponentially growing cells were transfected with 250 ng DNA (TcoClec or control) using Lipofectamine 2000 in OPTIMEM (Invitrogen) according to the manufacturer's instructions and processed for IFA 24 h post-transfection. To control transfection efficiency, the pcDNA3.1 Bilbo plasmid was used 22. A transfection without DNA was used as negative control.