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BIOL2262 GENETICS & MOLECULAR BIOLOGY ASSIGNMENT SHEET #2 -2013 Mutations To be submitted by mail or hardcopy to Reception, Level 1, Building

3, RMIT City Campus by 5.00pm on Friday, 24th May 2013. The submitted solutions must be hand written and accompanied by a signed RMIT submission cover sheet. 1. A strain of E. coli carries a substitution mutation causing the terminator codon 5-UAG-3 to occur at an interior position in a particular type of mRNA. This strain also carries a suppressor mutation that alters the anticodon of a type of tRNA molecule so that it reads 3-AUC-5. This type of tRNA reads this terminator codon and inserts the amino acid tyrosine. a. What effect would the presence of both of these mutations together have on the polypeptide translated from the mRNA carrying the mutant terminator codon?

The polypeptide would have a tyrosine at the triplet containing the mutant terminator codon. If the actual terminator codon is UAG, then the polypeptide would have a tyrosine at its actual terminator codon as well and extend until the next UAA or UGA triplet in the mutant mRNA. If the actual terminator codon is UAA or UGA, however, then the length of the polypeptide would be unaffected. b. Do you expect that there might be other genes whose translation would be altered by the presence of the tRNA produced by this suppressor gene? If so, describe the circumstances under which translation would be altered and explain how it would be altered. Genes with the terminator codon UAG would all be extended and have a tyrosine in place of its normal terminator codon. They would extend until the nearest UAA or UGA downstream from the normal terminator codon. 2. A gene in a haploid organism produces an mRNA transcript containing a section with three adjacent codons that are identical and that code for leucine. Sequencing mRNA from the same gene following exposure of the organism to a mutagenic chemical indicates that codons at these three sites now code for tryptophan, serine, and lucine. Assume that the mutations that occurred were single-base substitutions. What was the makeup of the codons present in the original mRNA?

Original 5-UUGUUGUUG-3 leu-leu-leu Mutant 5-UGGUCGUUG-3 trp-ser-leu 1


BIOL2262 GENETICS & MOLECULAR BIOLOGY Assignment Sheet #2 Mutations

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Human hemoglobin is a complex protein molecule made up of four polypeptides joined to an iron-containing heme group. In normal human adult hemoglobin, haemoglobin A, or HbA, two kinds of polypeptides designated as alpha and beta are found. Two identical alpha and two identical beta chains plus the heme group make up each molecule of hemoglobin A. Hemoglobin S, or HbS, is a haemoglobin variant occurring in individuals affected with the heritable disorder sickle cell anaemia. A comparison of the amino acid sequences of the polypeptides in haemoglobins A and S indicates that the alpha chains in the two molecules are identical, but that the beta chains differ in a single interior amino acid. a. How many kinds of genes are necessary for the synthesis of haemoglobin S?

Only one gene is necessary for the synthesis of hemoglobin S. b. Assuming that the gene for the beta chain of HbS arose from the gene for the beta chain of HbA through a single mutation, what general type of mutation was most likely responsible? A point mutation is most likely responsible. c. The difference between the beta chains of haemoglobin S and A is restricted to the sixth amino acid from the amino (-NH 2) end of the polypeptide. In haemoglobin S, the amino acid at the site is valine and in haemoglobin A it is glutamic acid. From your knowledge of the genetic code (and with reference to the genetic code) what can you deduce about the specific (and simplest) base alteration in the beta polypeptide that would produce the amino acid substitution. The simplest base alteration would be a substitution from an A to a U at the 2 nd position in the sixth triplet. There are two possible codons for glutamic acid (GAA and GAG) and a substitution from an A to a U at the 2 nd position would make both codons code for valine instead.

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BIOL2262 GENETICS & MOLECULAR BIOLOGY Assignment Sheet #2 Mutations