FIG. 2.
In vitro LM biosynthesis in M. smegmatis overexpressing Rv3779. Crude cell extracts (4 mg protein) of mc2155/pVV16 and mc2155/pVV16-Rv3779 were incubated with 1.0 μCi of GDP-[14C]Man (specific activity, 305 mCi/mmol) for 4 and 24 h. The reaction was stopped by adding CHCl3-CH3OH (2:1, vol/vol), and the LM/LAM contained in the cell pellet was extracted with hot phenol. LM/LAM separated on 10 to 20% Tricine gel and subsequently blotted to a nitrocellulose membrane was revealed by autoradiography. The leftmost column shows molecular mass markers (in kilodaltons).