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DNA and Protein Synthesis Proteins are vital to living organisms. They are involved in chemical reactions, oxygen transport, muscle contraction, sensory perception, blood clotting, and many other activities. The great variety of roles requires equal variety in the structure of protein molecules. This variety is achieved by means of 20 different amino acids linked end to end in polypeptide chains. The sequence of amino acids in a polypeptide chain determines the structure and biological role of a particular protein. This sequence is determined by two nucleic acids: DNA and RNA. Both of these molecules contain the “blue prints” for building proteins. In this lab, you will use these blue prints to build your own “proteins”. Procedure: Part A A. Obtain a cup of beads from the side counter and three of the lacing strings. The beads represent the amino acids, the string represents the peptide bonds that holds the amino acids together. B. Transcribe the DNA sequence into mRNA. C. Using the codon chart provided, translate the mRNA codons into a polypeptide chain. Find the correct color of bead and string them together. * NOTE* Make sure you do not start the process of translation until you reach a “Start” codon and stop at a “stop” codon. D. When you finish, come up the front and check your polypeptide (bead sequence) with the original to check for accuracy. DNA Sequence: CCAGGTATACCCGGCTTGACTAGGGGTCGCGAATACGACTAGGC mRNA Sequence: Circle codons starting at the “Start” codon and stop and the “Stop” codon. Amino Acid Color Sequence: Part B: Substitution Mutation A. You will now observe how a change in the sequence of a single DNA nucleotide affects protein synthesis. The DNA strand is the same as in Part A, except for a single substitution. Identify the substitution mutation and circle it. B. Transcribe the DNA sequence into mRNA. C. Using the codon chart provided, translate the mRNA codons into a polypeptide chain. Find the correct color of bead and string them together. * NOTE* Make sure you do not start the process of translation until you reach a “Start” codon and stop at a “stop” codon. DNA Sequence: CCAGGTATACCCGGCTTGATTAGGGGTCGCGAATACGACTAGGCT mRNA Sequence: Circle codons starting at the “Start” codon and stop and the “Stop” codon. Amino Acid Color Sequence: Part C: Frameshift Mutation A. You will now observe how a change in the sequence of a single DNA nucleotide affects protein synthesis. The DNA strand is the same as in Part A, except for a single Insertion of a nucleotide. Identify the insertion mutation and circle it. B. Transcribe the DNA sequence into mRNA. C. Using the codon chart provided, translate the mRNA codons into a polypeptide chain. Find the correct color of bead and string them together. * NOTE* Make sure you do not start the process of translation until you reach a “Start” codon and stop at a “stop” codon. DNA Sequence: CCAGGTATACCCCGGCTTGACTAGGGGTCGCGAATACGACTAGGCT mRNA Sequence: Circle codons starting at the “Start” codon and stop and the “Stop” codon. Amino Acid Color Sequence: A Yellow Yellow Yellow Yellow Light Blue Light Blue Light Blue Light Blue Green Green Green Green Dark Blue Dark Blue Dark Blue Dark Blue A G U C G Orange Orange Orange Orange Red Red Red Red Yellow Yellow Pink Pink Purple Purple Purple Purple A G U C Red Red Red Red Light Blue Light Blue Light Blue Light Blue A G U C A G U C Stop Codon Stop Codon Stop Codon Green U Pink Green Pink Green C Dark Blue Dark Blue Dark Blue Dark Blue Purple Purple Purple Purple Pink Pink Yellow Yellow Orange Orange Orange Orange A G U C Third Base in Code Word First Base in Code Word mRNA Codon Second Base in Code Word BE CAREFUL your message only starts with the translation at the START codon and you stop with the “stop” codon. Also remember to watch the direction of the DNA. Start codon = AUG Stop codons =UAA, UAG, UGA Analysis and Conclusions: 1. Which part of the simulation represented a polypeptide? 2. Which part of the simulation represented an amino acid? 3. What is a peptide bond, and how is it shown in the simulation? 4. What was the affect of the substitution mutation on the polypeptide chain? 5. What was the affect of the frameshift mutation on the polypeptide chain? 6. What is the process of transcription? 7. What is the process of translation? 8. What enzyme is responsible for transcription? 9. What is the job of a ribosome? 10. What is the job of tRNA? 11. Which part of this simulation would occur in the nucleus of a cell? 12. Which part of this simulation would occur in the cytoplasm of a cell? 13. Frameshift mutations are generally deemed as more harmful as substitution mutations. Why? 14. How is it possible for a substitution mutation to happen, but would have no effect on the amino acid sequence?