Identification of the genes and gene products involved in the biosynthesis of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine has lagged behind that in many other fields because of difficulties encountered in purifying the respective proteins. Nevertheless, most of these genes have now been identified. In this review article, we have highlighted important new findings on the individual enzymes and the corresponding genes of phosphatidylcholine synthesis via its two major biosynthetic pathways: the CDP-choline pathway and the methylation pathway. We also review recent studies on phosphatidylethanolamine biosynthesis by two pathways: the CDP-ethanolamine pathway, which is active in the endoplasmic reticulum, and the phosphatidylserine decarboxylase pathway, which operates in mitochondria. Finally, the two base-exchange enzymes, phosphatidylserine synthase-1 and phosphatidylserine synthase-2, that synthesize phosphatidylserine in mammalian cells are also discussed.