What s the Point? --- Point, Frameshift, Inversion, & Deletion Mutations
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1 What s the Point? --- Point, Frameshift, Inversion, & Deletion Mutations Introduction: In biology, mutations are changes to the base pair sequence of the genetic material of an organism. Mutations can be caused by copying errors in the genetic material during cell division, by exposure to ultraviolet or ionizing radiation, chemical mutagens, or viruses. In multicellular organisms, mutations can be subdivided into germ line mutations, which can be passed on to descendants, and somatic mutations, which cannot be transmitted to descendants in animals. Proteins are made of amino acids that are strung together in a chain. Each three-letter DNA sequence, or codon, encodes a specific amino acid. If a change in the DNA sequence occurs, the instructions for making the proteins will be changed producing a mutation. Types of Mutations: Substitution A substitution is a mutation that exchanges one base for another (i.e., a change in a single chemical letter such as switching an A to a G). 1
2 Insertion Insertions are mutations in which extra base pairs are inserted into a new place in the DNA. Deletion Deletions are mutations in which a section of DNA is lost, or deleted. Frameshift Since protein-coding DNA is divided into codons three bases long, insertions and deletions can alter a gene so that its message is no longer correctly read and translated. These changes are called frameshifts. Objective: Students will use DNA sequences and corresponding paper templates of mrna strands to show various mutations and the effect that changing these genes has on the construction of the amino acid sequence of proteins. Materials: DNA strand template mrna strand template Scissors Tape Crayons or colored pencils Small paper clips Computer with Internet Access Procedure (Part A) DNA Sequence & Proteins: 1. Color code the five genes on Template 1 as follows: Gene 1 red, Gene 2 blue, Gene 3 green, Gene 4 yellow, and Gene 5 violet. 2
3 2. Cut out the 5 genes and tape them together in order, 1 5, to assemble a chromosome with its DNA base sequence. This is your DNA template. 3. Record the sequence of the DNA bases for each gene in Table Use the base-pairing rule to complete the mrna nucleotide sequence that would read and copy the bases on each DNA gene in the DNA template you made. Write the correct letter for the base (a, T, C, or G) in each box of the mrna gene. REMEMBER: ADENINE (A) PAIRS WITH URACIL (U) AND CYTOSINE (C) WITH GUANINE (G) ON RNA. 5. After each base is written on the mrna genes, color code them as you did for the DNA genes; Gene 1 red, Gene 2 blue, Gene 3 green, Gene 4 yellow, and Gene 5 violet. 6. Do NOT tape these 5 mrna genes together. Instead, join them together in the correct order (1 5) with small paper clips so mutations can be made. This is your mrna template. 7. Record the sequence of the mrna bases for each gene in Table Ribosomes read the bases on mrna three at a time (called a codon) to assemble amino acids. Use the codon table to make a list of the correct sequence of amino acids that the ribosome would assemble when it reads the mrna template strand. Record this amino acid sequence in Table 1. 3
4 mrna Codon Table Procedure (Part B) - Modeling Mutations: Use the mrna template strand that you assembled and paper clipped together to simulate each of these types of mutations: 4
5 Substitution 1. The last base triplet of DNA Gene#1 is TTT which is copied by mrna as the codon AAA. (Refer to your data for Gene #1 on your Sequence Table.) What amino acid did this codon code for when it was read by the ribosome? 2. If the base guanine (G) is substituted for the final base thymine (T) on the DNA gene when DNA is replicated, write this new sequence for the DNA gene, mrna gene, and amino acid sequence in Data Table 2. Did this base substitution change the sequence of amino acids --- yes or no? 3. If the base substitution for this same base had been cytosine, not guanine, would the amino acid sequence have been different --- yes or no? Explain why this did or did not produce a change in the amino acid sequence. 4. NOW MAKE THIS MUTATION ON YOUR mrna STRAND AND THEN RE- CLIP THE STRAND TOGETHER. Deletion and Frameshift 1. The base sequence on DNA Gene #2 is --- GGG CCC TAT TAA GCA. 2. Write the mrna sequence and amino acid sequence for Gene #2 in Data Table If the last thymine (T) in the DNA gene is deleted, the new base sequence will be --- GGG CCC TAT AAG CA. This will change the mrna and amino acid sequence because it changes the reading frame by shifting it. Write the NEW sequences for mrna and amino acids in Data Table NOW MAKE THIS MUTATION ON YOUR mrna STRAND AND THEN RE- CLIP THE STRAND TOGETHER. 5
6 Insertion & Inversion 1. In order for a ribosome to know when to start and stop reading an mrna strand when assembling amino acids, specific start and stop codons appear at the beginning and end of the mrna strand. 2. AUG on mrna is the start codon called methionine, while UAA, UAG, and UGA do not code for an amino acid and signal the ribosome to stop. 3. Look at DNA Gene #5. What are the last 3 bases? What would be the mrna codon for these 3 bases? What does the codon stand for? 4. The sequence of the bases on DNA Gene #5 is --- CTG CAT CGG CGC ATT. 5. Insert and extra thymine base (T) in the 7 th position (after the sixth T and the next letter C). Write the new base sequence (16 letters total). New DNA base sequence: 6. Determine the mrna and amino acid sequence made from this changed DNA Gene #5. REMEMBER, IF A STOP CODON APPEARS ON mrna, NO MORE AMINO ACIDS WILL BE ASSEMBLED. New mrna Sequence: New Amino Acid Sequence: How did this gene mutation affect the building of the protein? 7. Inversion is a phenomenon in which a gene turns 180 o and attaches in reverse order. If Gene #5 inverts, what will be the first 3 bases on DNA? What is the mrna codon for these 3 bases? What do these codon stand for? What happens to the protein? 8. NOW MAKE THESE MUTATIONS ON YOUR mrna STRAND. 6
7 9. TAPE THE FINAL STRAND TOGETHER. Questions: 1. How many genes made up your DNA strand? 2. How many polypeptides would this DNA template code for? Using the following four codons, answer these questions. TAC GGT AAC CAT 3. What is the mrna transcript sequence? 4. What is the complementary DNA strand sequence? 5. If the 3 rd DNA base is deleted, what is the new mrna sequence? 6. If the base adenine is inserted in the 7 th position on the DNA strand, what is the new mrna sequence? 7. What is the amino acid sequence coded for by the original DNA strand? 8. What is the amino acid sequence after the deletion occurred? 9. What is the amino acid sequence after the insertion occurred? The nucleotide sequence for the left template strand of human hemoglobin is --- TAA TGT CGA CCG CTG GTC CAA GTC CTT TGA 10. Write the nucleotide sequence for the complementary DNA strand. 11. What would be the mrna transcript sequence? What is the amino acid sequence? 12. If the last two codons are inverted on DNA, what is the new mrna sequence and amino acid sequence? 7
8 Extension: Scientists working around the world have mapped the 46 human chromosomes. The project is called the Human Genome Project. Today, these mapped chromosomes with their genes can be studied to determine what genes may cause different characteristics or disorders in humans. One such mapping site is the National Center for Biotechnology s internet site, The Human Gene Map. Use this website to locate the chromosome and gene responsible for at least 5 different human genetic disorders. References: DNA Replication Animation Types of Mutations The Human Gene Map 8
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