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3-5 Biology 22 Problems in Gene Structure and Function 3. Four auxotrophic mutant strains of E. coli (1, 2, 3, 4) are each blocked at a different step of the same metabolic pathway. This pathway involves compounts W, X, Y and Z. Each of these compounds is tested for its ability to support growth of mutant strains, with the results given below. (+ = growth, -- = no growth). Compound Used as Supplement W X Y Z Mutant 1 + -+ + Mutant 2 + -+ -Mutant 3 + + + + Mutant 4 -- -+ -- a. Give the order of the compounds in the metabolic pathway and indicate the specific step that is blocked in each mutant strain. b. Several sets of double muations were produced. For example, double mutant 1,2 has mutant forms for both enzyme 1 and enzyme 2. Pairs of these double mutations were combined in partial diploids. Would a partial diploid consisting of each of the following pairs of mutations grow on minimal medium, without supplementation with W, X, Y and Z? 1. 1, 3 and 2, 4 2. 3, 4 and 2, 3 3. 1, 4 and 2, 3 4. Amino acids 1-5 were compared for an E. coli metabolic enzyme from a wild type strain and a strain with a double frameshift mutation having one nucleotide deletion followed by one nucleotide insertion. The results are shown below. Wild Type NH2- 1 MET 2 HIS 3 ALA 4 VAL 5 LYS -COOH Double NH2SER THR GLN LEU LYS -COOH Mutant a. Determine the base sequence of the mRNA for both the wild type and double mutant polypeptides. b. Using your wild type mRNA sequence, show which base was deleted and the site from which it was removed. c. Using your mutant sequence, show which base was added and where it was inserted. (There may be more than one possible solution.) 3-6 Biology 22 Problems in Gene Structure and Function 5. The sequence of eight amino acids of a protein from the bacterium E. coli is given below: 11 12 13 14 15 16 17 18 NH 2 Amino Acid TRP GLU THR SER VAL PHE ARG GLY -COOH Original Strain After exposing these bacteria to a mutagen, one mutant was isolated with different amino acids in this region of the protein. The mutation resulted from a change in a single nucleotide, either a substitution, insertion or deletion. What is the cause of the mutation? What is the sequence of codons for the original strain? Amino Acid MUTANT NH2- 11 GLY 12 ARG 13 HIS 14 PRO 15 SER 16 LEU 17 GLU 18 VAL -COOH 6. Nine mutants of the rII region in bacteriophage T4 DNA were analyzed for complementation. The results are given in the table below where + = complementation and -- = no complementation. Determine the number of cistrons represented by these mutations and indicate which mutations are found in each cistron. 1 2 3 4 5 6 7 8 9 1 -+ + + -+ + -+ 2 -+ -+ + -+ + 3 -+ + -+ + -4 -+ + -+ + 5 -+ + -+ 6 -+ + -7 -+ + 8 -+ 9 --