Analysis of metabolic control in vivo using molecular genetics

The aim of this brief review has been to illustrate the enormous power of molecular genetic techniques for testing out our, sometimes old, models of metabolic control. The technology allows us to ask some very simple but direct questions about the importance of specific enzymes in the control of flux. The answers, however, may not always be straightforward in that the absence of a change in pathway flux does not necessarily indicate that the enzyme is unimportant in flux control. Rather it would seem, from the relatively few studies that have been done, that the changes in the biological system which occur in response to a change in the concentration of a specific enzyme could tell us a lot about the role of that enzyme in metabolism and how it is controlled. I have restricted my discussion to studies in which relatively large numbers of cells have been modified genetically and subsequently characterized. This ignores a large and growing area in which micro-injection techniques are being used to introduce plasmid DNA or proteins into individual cells. The resultant phenotypes are then characterized at the single cell level using very sensitive optical techniques, such as fluorescence and bioluminescence. The reader is referred to a recent article for an example of this type of approach.