The two isogenic clones of L.g infected with, or depleted of LRV1 (termed LRV1+ or LRV1- respectively), used in this study were described and characterized previously [65 (link)]. Briefly, these lines were derived from the LRV1+ parent strain, L.g M4147 (MHOM/BR/75/M4147) or an LRV1-null derivative [66 (link)]. The parasites express similar levels of a firefly luciferase (5x107 photons/sec/106 parasites) from a LUC gene integrated stably into the small subunit gene of the ribosomal RNA locus.
Parasites were cultured in vitro as promastigotes at 26°C in freshly prepared Schneider’s insect medium (Sigma) supplemented with 10% heat-inactivated foetal bovine serum (PAA), 10mM HEPES and 50U/ml penicillin/streptomycin (Animed), 0.6 mg/L biopterin and 5 mg/L hemin (Sigma-Aldrich). Each passage yielded infectious metacyclic promastigotes after 6 days and stocks were kept for no longer than 5 passages.
Parasites were cultured in vitro as promastigotes at 26°C in freshly prepared Schneider’s insect medium (Sigma) supplemented with 10% heat-inactivated foetal bovine serum (PAA), 10mM HEPES and 50U/ml penicillin/streptomycin (Animed), 0.6 mg/L biopterin and 5 mg/L hemin (Sigma-Aldrich). Each passage yielded infectious metacyclic promastigotes after 6 days and stocks were kept for no longer than 5 passages.
Free full text: Click here
