The unfolding of a tRNA molecule has been measured as a function of temperature (data below). Use the equation ∆G= ∆G0 centigrade–m (Tempcentigrade) to calculate the free energy of folding at 0° ad 30°C. This can be done in Excel. Temp (C).              Fraction unfolded 40.                       0.240012 50                        0.5 60                        0.747475  70                         0.891304  80.                        0.955556

Beer's Law to determine Protein Concentration You have purified a recombinant form of the p53 protein from E. coli and determined the A280 to be1.35. Calculate the molar and mass concentration of the purified protein if the extinction coefficientand molecular weight of p53 is 35,410 M-1 cm-1 and 43,653 Da, respectively (l = 1 cm).

Explain how lysyl-tRNA synthetase protein in bacterial cells typically undergoes folding with necessary structures and steps, any ATP hydrolysis, cap structure, etc.

Explain why each of the criteria listed below are important in selection of a guide RNA (hint: look under Instructions on CHOPCHOP).  Under what conditions do these criteria decrease the efficiency of binding of guide RNA to its target sequence (e.g. does more, or less, self-complementarity decrease efficiency)? a) % GC (= # of G and C’s in the guide RNA sequence divided by 20 – the length of the guide RNA; this may also be called GC content) b) Self-complementarity: c) Off-Targets (0, 1, 2, 3). In your answer, explain what the numbers 0, 1, 2, 3 indicate.  For example - Is it better to have off-target sites or no off-target sites? Are off-targets in the 1 column more or less likely to decrease efficiency than off-targets in the 3 column?

In studies using repeating copolymers, AC . . . incorporates threonine and histidine, and CAACAA . . . incorporates glutamine, asparagine, and threonine. What triplet code can definitely be assigned to threonine?

Which amino acid would be attached to a tRNA that read "GGU"?

Analyse and interpret the SCAM data in Fig. 1, and make a table (remember to include a legend) to indicate where each amino acid is located (cytoplasm, membrane, periplasm).