Objectives – Translation Part II 1. Describe the entire process of translation in prokaryotes. Be sure to include all necessary components, including the specific roles of IF’s, EF’s and RF’s. 2. How does the initiator tRNA differ from all other tRNA’s in translation? 3. What is the total energy expenditure required to make a given protein (lets say the protein is 100 amino acids in length)? 4. How does translation differ between prokaryotes and eukaryotes? 5. Where is the enzyme peptidyl transferase located? 6. Describe the “fragment assay” of Monro et al. What were the conclusions of these experiments? Understand this assay and be able to interpret alternative results. 7. How did the “reconstitution” studies of the early 1980’s differ from Monro’s fragment assay? What were the results of these experiments? 8. Describe the “deprotienation” experiments of Noller et al. What were the results of this line of experiments? Understand this assay and be able to interpret alternative results. 9. Define codon. 10. What is the meaning of a degenerate code? 11. Why is the code considered nearly universal? 12. Why is the code considered non-overlapping? 13. Describe the cell free protein synthesis reaction. 14. How did Nirenberg and Matthaei utilize this assay in determining what certain codons coded for? 15. Explain the work of Lengyel and Speyer (random co-polymers in a cell free protein synthesis reaction). How did their work help in determining the genetic code? Be sure you understand and are able to interpret the results of this type of experiment. 16. In the early 1960’s Khorana was able to improve RNA synthesis. Describe his experiments and how they contributed to cracking the genetic code. 17. Describe the triplet binding assay of Nirenberg and Leder. Understand these experiments and be able to interpret results.