Genetic manipulation of Leishmania donovani threonyl tRNA synthetase facilitates its exploration as a potential therapeutic target
Fig 4
Characterization of genetically modified parasites.
(A) Aminoacylation activity of LdThrRS in the cell lysates of L. donovani WT, heterozygous (ThrRS/NEO) and rescue mutant (ThrRS/NEO[pThrRS+]) parasites. (B) Comparison of the growth curve characteristics of WT, heterozygous (ThrRS/NEO) and rescue mutant (ThrRS/NEO[pThrRS+]) promastigotes in M199 media. The experiment was repeated thrice in triplicate. The data shown here is from one experiment. (C) and (D) Comparison of the infectivity (C) and parasite load (D) of L. donovani WT, ThrRS/NEO and ThrRS/NEO[pThrRS+] parasites in J774A.1 murine macrophage cell line. The stationary phase promastigotes were used to infect murine macrophage cell line J774A.1 at an MOI of 20:1. After 48 h of infection, cells were stained, and amastigotes were counted visually. The results signify mean ± S.D with n = 3, *P < 0.05, **P < 0.01 statistical difference from the wild-type control.