Analysis of lipid biosynthesis and location.
A procedure for metabolic labeling of all cellular lipids starting with a culture of mycobacteria is described in this chapter using either a pulse-chase or a simple labeling experimental design. Three fractions are produced for subsequent lipid analysis: (1) the culture filtrate; (2) a readily released surface lipid fraction; and (3) the killed, labeled bacteria. A standardized, TLC-based method for general lipid analysis that can be used to quantify the labeling of all the mycobacterial lipids is given as well as a protocol for analyzing the fatty acyl moieties of the lipids.
Wheeler
Paul Robert
PR
Tuberculosis Research Group, Veterinary Laboratories Agency (Weybridge), New Haw, Addlestone, Surrey KT15 3NB, UK. p.wheeler@vla.defra.gsi.gov.uk
D002855:Chromatography, Thin Layer / Q000379:methods*; D005227:Fatty Acids / Q000032:analysis / Q000378:metabolism; D050356:Lipid Metabolism; D008055:Lipids / Q000032:analysis*; D008967:Molecular Biology / Q000379:methods; D009161:Mycobacterium / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods
D005227:Fatty Acids; D008055:Lipids