Figure 1
(A) Schematic representation of L-methionine biosynthetic pathway in M. tuberculosis. The enzymes involved in L-methionine biosynthetic pathway of M. tuberculosis are shown. Rv3341, highlighted in the box encodes for homoserine acetyl transferase that catalyses the first committed step in L-methionine biosynthesis. (B) Silencing of Rv3341 expression in M. tuberculosis using CRISPRi approach. The effect of CRISPRi on metX expression in knock down strain was quantified by qPCR using gene specific primers. The data shown in this panel is mean ± S.E. fold change of ΔCT obtained from two experiments performed in duplicates (C,D) Effect of metX repression on M. tuberculosis growth in vitro. The effect of conditional repression of metX on in vitro growth of M. tuberculosis was determined by measuring absorbance (OD600nm, C) at regular intervals. In addition, 10.0 fold serial dilutions of cultures were spotted on MB7H11 plates (D). The plates were incubated at 37 °C for 3–4 weeks. The data shown in panel C is mean ± S.E. of OD600nm values obtained from three independent experiments. The data shown in panel D is representative of two independent experiments.
