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Name ________________________________ Lab: Nicotine Addiction Nicotine is a substance that acts as a stimulant in mammals and is the main factor responsible for the dependence-forming properties of tobacco smoking. According to the American Heart Association, nicotine addiction has historically been one of the hardest addictions to break. Scientists say that they have pinpointed a genetic link that makes people more likely to get hooked on nicotine, causing them to smoke more cigarettes, making it harder to quit, and leading more often to lung cancer. In this lab, we will refer to this genetic link as the fictitious lung cancer predisposition (LCP) gene. Is this molecule organic or inorganic? Pause for the Cause: Vocabulary allele ______________________________________________________________ chromosome _________________________________________________________ DNA ______________________________________________________________ gene _______________________________________________________________ genome ____________________________________________________________ genotype ___________________________________________________________ phenotype __________________________________________________________ predisposition ________________________________________________________ trait ______________________________________________________________ Part 1: Pedigree Investigation In this part of the lab, we will explore a pedigree chart to see how it can be applied to a realworld situation. We will use the following website to fill in the pedigree chart on the back of this page: http://learn.genetics.utah.edu/content/addiction/genetics/pi.html 1 g-$. Wilson Commencement Academy Rochester, NY 2 g-$. Wilson Commencement Academy Rochester, NY Once you’ve filled in the pedigree, answer the questions below in complete sentences on a separate sheet of paper. 1. 2. 3. 4. 5. 6. 7. What is a pedigree? What is the value of pedigrees for researchers studying human disease? What is a risk factor? Why do researchers consider environmental risk factors when studying a trait? What were some of the risk factors for nicotine addiction? What clues in a pedigree would indicate that a trait can be inherited? Why is it difficult to predict whether or not a disease like nicotine addiction will be an issue for an individual? 8. If a pedigree shows evidence that a trait can be inherited, what might scientists do next? Part 2: Research Review Now read through the research article that your teacher has given you. Once you are done reading, answer the questions below in complete sentences on a separate sheet of paper. 9. According to the research, what is determined by the environment? 10. According to the research, what is determined by genes? Part 3: Punnett Square Predictions In this section of the lab, you will make predictions using Punnett squares. You will then use pennies to simulate the crosses. Finally, you will compare the actual percentages with the predicted percentages. The trait that you are investigating– lung cancer predisposition– is coded for by a gene that gives someone a predisposition to nicotine addiction and, potentially, lung cancer (the LCP gene). T represents the dominant allele (no LCP); t is the recessive allele (LCP). 11. Fill in the missing genotype names and phenotypes for the following possibilities: Genotype Genotype Name Phenotype If this person smokes… TT homozygous dominant ________ __________________ Tt ________________ no LCP __________________ t t ________________ ________ __________________ 3 g-$. Wilson Commencement Academy Rochester, NY 12. Use a Punnett square to predict the probability of offspring in a cross where the parents are both Tt. This Punnett square is set up for you below. What proportion of the offspring (out of 4) will have: No LCP ______ LCP ______ Note: These are your predicted ratios. Now you will determine the actual percentages by using pennies to represent the crosses. You have two pennies. On one side of the penny is the letter T (representing the dominant allele) and on the other side is the letter t (representing the recessive allele). This penny represents a parent that has the hybrid a.k.a. heterozygous genotype Tt. A second penny, labeled the same, represents the other parent. One partner is going to play the role of the female and the other will play the role of the male. Very romantic! When the coin is flipped, you are determining what haploid sperm or what haploid egg is being donated to the diploid zygote (fertilized egg cell). 13. Flip the two pennies to find the genotype of your offspring. Did you get TT, Tt, or tt? ____ 14. What is the phenotype of your offspring: predisposed to lung cancer or not? ___________ 15. To determine actual percentages, you will flip your coins 50 times, recording in the table below how often each combination comes up. Use tally marks to record your data and then summarize as a percentage. Actual Ratios Genotype (alleles) Tally Total homozygous dominant ( TT ) hybrid a.k.a. heterozygous ( __ __ ) homozygous recessive ( __ __ ) 4 g-$. Wilson Commencement Academy Rochester, NY Total X 2= ____% 16. Now combine the data from the previous page to calculate phenotype percentages. Phenotype Total % No LCP (add TT + Tt ) LCP (tt) Your totals above represent a percentage. Your proportions from the Punnett square in your prediction can also represent a percentage. 1/4 = 25 % 2/4 = 50% 3/4= 75% 4/4 = 100% 17. Now compare your predicted percentages to your actual percentages in the chart below. Predicted %s (from the square you did) Actual %s (from the flips) TT Tt tt No LCP LCP 18. When you compare the predicted values to the actual values, are they the same, close to the same, or not at all the same? _______________________________________________ ***You will now repeat the procedure for parents that are Tt and tt.*** 19. First, make your predictions by setting up a Punnett square for the parents. This one is not set up for you. Sick! What proportion of the offspring (out of 4) will have: No LCP ______ LCP ______ 5 g-$. Wilson Commencement Academy Rochester, NY ***Replace one of your pennies with a tt penny.*** 20. Perform the flips with your new set of parents. Record your data in the table below. Tally Total Total X 2= ____% Tt tt 21. What percentage of your offspring are Tt? ___ What percentage do not have LCP? ___ 22. What percentage of your offspring are tt? ___ What percentage do have LCP? ___ 23. Compare the predicted percentages of the cross to the actual percentages. Predicted %s (from square) Actual %s (from flips) No LCP LCP Again, answer the following in complete sentences on a separate sheet of paper: 24. 25. 26. 27. Why are the predicted percents rarely the same as the actual percents? Why are Punnett squares useful for determining phenotype chances in offspring? What do the pennies represent in the simulation? When you toss the coin to see which side lands up, you are actually simulating what part of the process of sexual reproduction? 28. When you put the two coins that are flipped together, you are simulating what part of the process of sexual reproduction? 29. Can you accurately determine a person’s genotype by observing their phenotype? Explain. 30. If you are genetically predisposed to nicotine addiction because you have the LCP gene, what choices can you make to avoid being addicted to nicotine and, therefore, dying from lung cancer? 6 g-$. Wilson Commencement Academy Rochester, NY Part 4: Mapping the LCP Gene Now that we have some idea of how this LCP gene can be inherited, we want to locate it. Scientists currently think that there are about 23,000 protein-coding genes in the human genome spread out over our 46 chromosomes. Somewhere, in this human genome, is our LCP gene. The human karyotype shows all of the chromosomes that we have in the nuclei of our body cells. Each of our body cells has this same set of genetic information— the same genome. In the space below, perform the following tasks: 31. 32. 33. 34. Label the nuclear membrane. Draw the human karyotype. Label each chromosome pair with a number. For the last chromosome pair– the sex chromosomes– label whether you are XX or XY. Separate sheet: 35. What percentage of these chromosomes came from each parent? 36. What process made it possible for your parents to donate only half of their chromosomes to you in their haploid gametes? 37. Deep thoughts: If males have XY as their sex chromosomes and females have XX, who determines the gender of the offspring? 38. On average, how many genes are found on each chromosome? 7 g-$. Wilson Commencement Academy Rochester, NY Now we will zoom in on one pair of chromosomes where the LCP gene is found— pair 19. Because this pair of chromosomes code for the same genes, they are called homologous chromosomes (“homo” means the same). Because one of the chromosomes is inherited from the organism’s mother and the other from the father, homologous chromosomes are rarely identical. We have enlarged an image of chromosome pair 19 below. Your task is to consider the traits that you and your partner have and predict what the gene map of your future offspring could look like. This one is from the dad! This one is from the mom! 39. Draw the LCP gene as a line 2.8 cm up from the bottom. Give you or your partner the dominant “T” allele with an orange line if nicotine addiction does not seem to run in the family. Otherwise, draw a red line for the recessive “t” allele if nicotine addiction does run in the family. 40. Draw the tongue roller gene as a line 1.7 cm up from the bottom. Give you or your partner the dominant “R” allele with a blue line if the tongue can curl or a recessive “r” allele with a green line if the tongue can’t roll. 41. Draw the attached earlobe gene as a line 4.2 cm up from the bottom. Give you or your partner the dominant “A” allele with a purple line if the earlobes are attached to the head or a recessive “a” allele with a brown line if the earlobes are swaying in the wind. Separate sheet: 42. What is the relationship between chromosomes, genes, and the nucleus? 43. What is a gene map? 8 g-$. Wilson Commencement Academy Rochester, NY Part 5: Making the LCP Protein Now that we have located the LCP gene on chromosome pair 19, we can splice that gene out of the genome with the help of a specific restriction enzyme and look at its code. Then we can see how this code forms an actual protein. Here’s an analogy: DNA is like a recipe, two-dimensional words on paper. Proteins, on the other hand, are like the delicious three-dimensional macaroni and cheese, all bubbly and fantastic as it comes out of the oven. In the same way, you are three-dimensional, bubbly and fantastic…but without your DNA “recipe”, you wouldn’t exist. You feel me? Anyways, let’s see what this LCP gene looks like and what kind of protein it makes. When spliced from the genome, the LCP gene has this sequence of DNA bases: TACAAACAGTTAGTCGTAGACACACCCTCAGTGGAT ___________________________________________ CAGGTCCGGGATATAAACCAAACGCCGCTCTCTCCCAAG ___________________________________________ AAAATGATGGGGTTTCGTCCATAACACCTTGTCACA ___________________________________________ ACAGCAAGACAAACAAGCAACATGGTTAACCTCTTA ___________________________________________ ATAACATTGATC ___________________________________________ 9 g-$. Wilson Commencement Academy Rochester, NY 44. The first step in making a 3-D LCP protein from the 2-D LCP gene “recipe” is transcription. During transcription, DNA is transcribed, or rewritten, into mRNA. Transcribe the DNA bases on the previous page into mRNA codons with the help of your teacher. This is very fun! 45. Now that you have the mRNA codons, you can translate them into the corresponding amino acid using The Universal Genetic Code chart provided by your teacher. This is also very fun! The first three letters of each amino acid should be used as an abbreviation on the next page. As you can see, amino acids are the subunits of proteins. Here is an easier chart for you to use after you have mastered the other one: 10 g-$. Wilson Commencement Academy Rochester, NY The LCP Protein 11 g-$. Wilson Commencement Academy Rochester, NY Use complete sentences on a separate sheet. 46. Infer where in the cell transcription takes place. 47. Use the following words to describe how proteins are made: DNA, mRNA, nucleus, protein, ribosome, transcription, translation. 48. Proteins are very specific shapes with very specific jobs. Draw what your LCP protein looks like and what it might look like if it was denatured by high heat or the wrong pH. Part 6: Modeling Genetic Organization Your teacher will now provide you with the following materials: a paper clip, some yarn, a large plastic bag, a small plastic bag, and a popsicle stick. Using what you know about genetic organization, model the connection between cells, nuclei, chromosomes, DNA, and genes. 49. When you are ready to explain what you came up with, call your teacher over and have him/her initial in this box if your model and explanation are correct: Use complete sentences on a separate sheet. 50. Use the following words to explain how genetic information is organized: cells, nuclei, chromosomes, DNA, and genes. Part 7: Treating the Recessive Trait Spend some time reading the passages, provided by your teacher, that relate to genetic engineering and the treatment of inherited disease. Use complete sentences on a separate sheet. 51. Describe one possible way that genetic engineering could help someone who has inherited a recessive LCP allele from both parents. 52. Describe another possible way that genetic engineering could help someone who has inherited a recessive LCP allele from both parents. You are done! Take a deep breath and relax! 12 g-$. Wilson Commencement Academy Rochester, NY 13 g-$. Wilson Commencement Academy Rochester, NY