Population Genetics and Evolution Lab 8 By: Milap Lavani Purpose The purpose of this lab is to understand the

Hardy-Weinberg law of equilibrium by being able to implement the Hardy-Weinberg to calculate the frequencies of alleles and genotypes in a population. In this lab we will also evaluate the evolutions of a population and its changes in allele frequency. EXERCISE 8A: ESTIMATING ALLELE FREQUENCIES FOR A SPECIFIC TRAIT WITHIN A SAMPLE POPULATION. Procedures: 1. Begin by placing a piece of the PTC test paper in your mouth. 2. Individuals who are able to sense the bitter taste, are considers to be tasters. 3. The frequency of tasters (p2 +2pq) is acquired by dividing the total number of tasters by the total number of students in the class 4. The frequency of nontasters (q2) is acquired by dividing the number of nontasters by the total number of students. 5. Use the calculated values and place them in the table 8a. 6. To calculate the frequencies of p and q, use the Hardy-Weinberg equation. Q can be found by taking the square root of q2. p can be found by subtracting 1 by q (1-q=p). 7. Use the calculated values and place them in the table 8a Hypothesis: If the numbers of PTC tasters are determined within the population of the class, then using the Hardy-Weinberg equation one will be able to determine the allele frequencies of the class. Independent variable: Amount of PTC tasters Dependent variable: allele frequencies of the class Constants: type of PTC paper Questions 1) What is the percentage of Heterozygous tasters (2pq) in your class?__________ 2) What percentage of the North American populations is heterozygous for the taster trait? 49.5%

Data Tables: Table 8a: Phenotypic Proportions of Tasters and Non-Tasters and Frequencies of the Determining Alleles Allele frequency based on the H-W Phenotype Equation Tasters (p=2pq) Class Population North American Population # % Non-tasters (q) # % p q

0.55

0.45

EXERCISE 8B CASE STUDIES Case 1-A Test of an Ideal Hardy-Weinberg Population. Procedure: 1. Each person should have an initial set of alleles of A,A,a,a. Each allele should be written on its own card. 2. After shuffling the cards, turn over the one card which will represent the allele that you have passed on to your offspring. 3. Your partner should do the same, thus having two alleles for your offspring. 4. Each student pair should produce two offspring; therefore, the cards should be shuffled before creating another offspring. 5. Write down the genotype (alleles) for one of the offspring on the data page, while your partner writes down genotype of the second offspring. 6. You and your partner now become the next generation, by assuming the genotype of the offspring that you are designated to. 7. Next, the students should obtain the cards required to assume their new genotype. 8. Randomly seek another student to mate with to produce the next set of offspring. Repeat steps 2-6 until 5 generations of offspring have been produced. 9. The teacher will collect the classs data after the 5 generation. 10. Calculate the allele frequencies
Number of A Alleles present at the fifth generation. Number of offspring with genotype AA__________________X 2=__________________ A alleles Number of offspring with genotype Aa__________________X 1= __________________A alleles Total=__________________A alleles

p=

_________Total number of A alleles_________ Total number of alleles in the population (number of student X2)

________

Number of offspring with genotype aa__________________X 2=__________________ a alleles Number of offspring with genotype Aa__________________X 1= __________________a alleles Total=__________________a alleles

q=

_________Total number of a alleles_________ Total number of alleles in the population (number of student X2)

________

Hypothesis: If the experiment is conducted under ideal conditions, then we will be able to observe a change in allele frequency of an ideal Hardy Weinberg population. Independent variable: Alleles (A,A,a,a) Dependent variable: Allele frequency in generation of 5 Constants: same alleles Questions: 1) What does the Hardy Weinberg equation predict for the new p and q? a. The new p and q predicts the frequency of AA alleles to be 51% while allele aa to be 24%. 2) Do the results you obtained in this simulation agree? If not, why not? a. The results that I obtained in this simulation generally agree because the sum of p and q equals 1. Also the frequency of the dominate allele and the recessive allele are almost equivalent, and allele Aa is the largest because it carries the most alleles. 3) What major assumptions were not strictly followed in this simulation? a. Natural factors that could cause a change in breading was not included into this simulation. Class II-Selection Procedure: 1. Similar to case I start with your initial genotype and produce fertile offspring. Except every offspring with the genotype aa dies. 2. The parents will continue to produce until a fertile offspring is created. Record your generations alleles in the data page, and calculate the new p and q.

Hypothesis: If the offspring with alleles aa die, then according to the Hardy-Weinberg law genotype frequency will increase for alleles Aa, and AA, however genotype aa will cease to exist. Independent variable: Alleles (A,A,a,a), however every aa offspring dies. Dependent variable: Allele frequency in generation of 5 Constants: same alleles Questions: 1) How do the new frequencies of p and q compare to the initial frequencies in Case I? a. Through natural selection offsprings with genotype aa were rejected, thus leaving only offspring with the alleles AA or Aa. Therefore, the p frequency in this case was higher than case I, and the q frequency is lower than those of case I 2) What major assumptions were not followed in this simulation a. The assumption that homozygous recessive will die 100% of the time and that homozygous dominant will survive 100% of the time violates natural selection because the parents continue to mate until a dominant phenotype is born. 3) Predict what would happen to the frequencies of p and q if you simulated another five generations. a. The frequency of the dominant allele (q) will continue to increase, while the frequency of the recessive allele (p) decreases. However, p will never be 0 because heterogeneous (Aa) will remain. 4) In a large population would it be possible to completely eliminate a deleterious recessive allele? Explain. a. It is impossible to completely eliminate a deleterious recessive allele; though people who are homozygous recessive will die before they could produce offspring, carriers will be able to live fit lives and still pass on this gene. Class III-Heterozygote Advantage Procedure: 1. Similar to case I start with your initial genotype and produce fertile offspring. 2. Except, every offspring with the genotype aa dies, and every genotype of AA has a 50% chance of survival. This is determined by flipping a coin, tails it lives and heads it dies. 3. The parents will continue to produce until a fertile offspring is created. Record your generations alleles in the data page, and calculate the new p and q 4. After 5 generations the teacher will collect the genotypes of the class. 5. Then calculate the p and q frequencies, and place it in the data page. 6. If time permits then proceed to the next 5 generations.

Hypothesis: If the offspring with alleles aa dies and alleles AA have a 50% of survival, then according to the Hardy-Weinberg law, genotype frequency will increase for alleles Aa, however genotype aa will cease to exist and allele AA will have a decreased frequency compared to its ideal. Independent variable: Alleles (A,A,a,a), however every aa offspring dies and alleles AA have a 50% of survival Dependent variable: Allele frequency in generation of 5 Constants: same alleles Questions: 1) Explain how the changes in p and q frequencies in Case II compare with Case I and Case III. a. The frequency of p in cases II was higher than that of case I because only genotype AA and Aa survived. Therefore, the frequency of q was lower in case II when compared to case I because genotype aa died. The frequency of p in case III has decreased drastically when compared to case II because genotype AA only survives 50% of the time. However, in both Case II and case III genotype aa does not survive. 2) Do you think the recessive allele will be completely eliminated in either Case II or Case III? a. The recessive allele will never be eliminated because there will always be heterozygotes 3) What is the importance of heterozygotes, the heterozygote advantage, in maintaining genetic variation in populations? a. Heterozygotes have both alleles, consequently they are required for genetic variation.

Class IV-Genetic Drift Procedure: 1. Divide the lab into several smaller populations. Thus interaction could only occur within the group. 2. Then go thought the five generations as you did in case I. 3. Record the new genotypic frequencies and calculate the new p and q frequencies. Hypothesis: If genetic drift occurs, then there will be a dominate allele with in the population. Independent variable: genetic drift Dependent variable: Allele frequency in generation of 5

Constants: same alleles Questions: 1) Explain how the initial genotypic frequencies of the populations compare. a. All groups had their initial genotypic frequencies as 100% Aa 2) What do your results indicate about the importance of population size as an evolutionary force? a. The smaller the population the greater the variance between allele frequencies because there are less options to mate with.

Hardy-Weinberg Equation Problems 1. In Drosophila the allele for normal length wings is dominant over the allele for vestigial wings (vestigial wings are stubby little curls that cannot be used for flight). In a population of 1000 individuals, 360 show the recessive phenotype. How many individuals would you expect to be homozygous dominant for this trait? a. 160 individuals are homozygous dominant and 480 are heterozygous. 2. The allele for unattached earlobes in dominant over the allele for the attached earlobes. In a population of 500 individuals, 25% show the recessive phenotype. How many individuals would you expect to be homozygous dominant and heterozygous for this trait? a. 125 are homozygous dominant and 150 are heterozygous 3. The allele for the hair pattern called Widows peak is dominant over the allele for no widows peak. In a population of 1000 individuals, 510 show the dominant phenotype. How many individuals would you expect of each of the possible genotypes for this trait? a. 510 are homozygous dominant, 410 are heterozygous, and 80 are homozygous recessive. 4. In the U.S. about 16% of the population is Rh negative. The allele for negative is recessive to the allele for Rh positive. If the student population of a high school in the U.S. is 2000, how many students would you expect of the three possible genotypes? a. 720 are homozygous dominant, 960 are heterozygous, and 320 are homozygous recessive. 5. In certain African countries, 4 percent of the newborn babies have sickle cell anemia, which is a recessive trait. Out of a random population of 1,000 newborn babies, how many would you expect for each of the three possible genotypes?

a. 640 are homozygous dominant, 320 are heterozygous, and 40 are homozygous recessive. 6. In a certain population, the dominant phenotype of a certain trait occurs 91 percent of the time. What is the frequency of the dominant allele? a. The frequency of the dominant allele is .91

Explanations of Results The frequency of p and q in a gene pool was evaluated in multiple scenarios tested in H-Ws ideal environment, a heterozygote advantage, natural selection, and genetic drift. In a H-Ws ideal environment pq. In the heterozygote advantage scenarios P increased while q decreased. In a natural selection both p and q decreased while 2pq remained high. In the genetic drift scenarios genotypic variation was low. Conclusion As initially stated by the hypotheses, It can be concluded that allele frequencies remain at equilibrium when there is an ideal environment like Case I. It has also been observed that when homozygotes die for a specific allele, it causes unbalance in the genotype frequencies. when individuals of a population breed with the same mate for several generations genetic drift can take place and genetic variance severely decreases. Only possible errors that may exist in the results can only be due to calculation errors, or procedural errors. The data acquired was predictable and there was no ambiguous data.