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Obtaining the eIF-5A gene knockdown parasites

XL Xinchao Liu
CL Chunjing Li
XL Xiaoyu Li
ME Muhammad Ehsan
ML Mingmin Lu
KL Ke Li
LX Lixin Xu
RY Ruofeng Yan
XS Xiaokai Song
XL XiangRui Li
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As eIF-5A is an essential constituent of cells and yeast, knockdown of eIF-5A provides an alternative method to continue the study. Three small interfering RNA (siRNA) targeting TgeIF-5A were designed and synthesized by Invitrogen (Shanghai, China) (Additional file 3: Table S3). Aliquots of 1 μM, 2 μM and 4 μM of each TgeIF-5A siRNA and negative control (stealth siRNA Negative Control Lo GC, Invitrogen) were transfected into 1 × 107 tachyzoites using the Gene Pulser Xcell™ Electroporation System (Bio-Rad, USA) at settings of 50Ω, 1500 V, and 25 µF. Parasites were added to the monolayer HFF cells after transfection and monitored to determine the transfection efficiency at 24 h after electroporation by real-time PCR and Western blotting (Additional file 6: Method S1 and Additional file 7: Method S2).

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