The bacterial one-hybrid assay holds E. coli cells hostage until our protein of interest can active transcription and translation of enzymes from the HIS3 and URA3 genes. These enzymes are needed to produce Histidine and Uridine, which are both needed for cells to grow and divide. Make an estimate, based on enzyme kinetics and any other information you may need, of how many HIS3 and URA3 enzymes will an E. coli cell need to grow and divide at a normal rate. How many are needed when we add 10 mM 3-amino-1,2,4-triazole (3AT) to the growth medium as a competitive inhibitor of HIS3?
Report your strategy by first listing all variables/ratios/numbers (with units) to be used and informally recognizing sources (copy-paste a link or “Table 4-1 in textbook” or “random number generator”). State all assumptions. For example, when estimating the amount of ATP used to do the world record bench press, we assumed that 100% of the energy from ATP hydrolysis went into the lift. It may have been more appropriate to assume perhaps 50% (intuitive guess).
Group variables used for unit conversions together and state starting and ending values. (Example: “Variables in List 1 were used to convert 3,600 Joules to 44 g of ATP”)
For equations, list forms from textbook and state how they were manipulated but don’t show steps. Use Insert Equation. Example: Solved