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The Study of X Linked Traits in the First and Second Generation of White Eyed Male and Wild Type Female Cross in Drosophila Christopher Ellis, Brad Brown, Tim S., Jenna McClure March 4 2011 ± May 6 2011 Honors Biology 10 Period 1
X LINKED TRAITS OF WHITE EYED MALE AND WILD TYPE FEMALE CROSS 2 Abstract .
the pupa ecloses into the adult stage. and have termed these exceptions Non-Mendelian Genetics. In the pupa stage. The short life span of fruit flies makes them an ideal subject for the study of genetics. as multiple generations can be studied in a short period of time. However. From egg to adult takes approximately 10 days at room temperature (25°C) (Arnini 1996). Females. were chosen for the study of X-Linked traits for multiple reasons. since Mendel¶s discoveries in genetics. Fruit flies are small insects approximately 3mm long. Drosophila. the life cycle of fruit flies is quite short. Life cycles of flies may be altered based on environment or certain genetic mutations.X LINKED TRAITS OF WHITE EYED MALE AND WILD TYPE FEMALE CROSS 3 Introduction Genetics is a topic that has been studied for hundreds of years. The diet of fruit flies is simple and does not require extraneous foods or materials (Manning 2008). the pupal case forms. have a pointed abdomen and are often lighter than males . or fruit flies. One of the most notable geneticists was Gregor Mendel. Additionally. Mendel determined that the offspring of two parents contains one gene from each parent (McClean 2000). lasting approximately 26 days for a female and approximately 33 days for a male (Arnini 1996). One type of Non-Mendelian genetic inheritance pattern is X-Linked traits. The life cycle of a fruit fly starts when an impregnated female fruit fly lays an egg. darkens and hardens for 4-6 days. Mendel studied basic inheritance patterns and gene expression using pea plants. other scientists have found exceptions to Mendel¶s rules. Fruit flies lay only one egg at a time. Males fruit flies have a smaller body with a rounded black tip at the end of their body. Finally. The egg hatches in 22 hours. and thus use minimal space when compared to other species. and the larvae eats and grows for four days. The larvae goes through three larva stages before entering the pupa stage. on the other hand.
meaning that it is carried on the X chromosome. wild type to white eyed. with half of them being male and half of the being female. The Punnett Square for the first generation can be found in figure one. wild type to white eyed. with half of them being male and half of the being female. and is recessive to wild type. with half of them being wild type female. wild type to white eyed. The purpose of the experiment was to determine the type of inheritance of the white eye genetic mutation within fruit flies. . wild type to white eyed. then the phenotypic ratio of the first generation will not be 4:0. one fourth of them being wild type male. which determines sex. and one fourth of them being white eyed male. and the phenotypic ratio of the second generation will be 3:1. Null Hypothesis If white eyed male fruit flies were crossed with wild type female fruit flies and the white eyed mutation is an X Linked Recessive trait. then the phenotypic ratio of the first generation will be 4:0. one fourth of them being wild type male.X LINKED TRAITS OF WHITE EYED MALE AND WILD TYPE FEMALE CROSS 4 (Arnani 1996). and the phenotypic ratio of the second generation will not be 3:1. Rationale Previous research has shown that the white eye mutation is an X Linked recessive trait. and one fourth of them being white eyed male. Hypothesis If white eyed male fruit flies were crossed with wild type female fruit flies and the white eyed mutation is an X Linked Recessive trait. with half of them being wild type female. By performing a Punnett Square for the first generation. the expected genotypic and phenotypic ratios can be found.
Because all of the females will have the same genotype. half female. and all of the males will have the same genotype.X LINKED TRAITS OF WHITE EYED MALE AND WILD TYPE FEMALE CROSS 5 XWt XWt = Wild Type Male XW Y = White Eyed Female Figure 1: Punnett Square for first generation White Eyed Male and Wild Type Female Cross Parent Generation: XWt XWt x XW Y Wild Type XWt White Eye XW White Eye Y XWt XW XWt Y Wild Type XWt XWt XW XWt Y This Punnett Square demonstrates that the expected phenotypic ratio for the first generation would be all wild type. Using this information a second Punnett Square for the second generation can be generated. The Punnett Square for the second generation can be found in figure two. only one Punnett Square needs to be created because there is no variance within the flies. half male. Figure 2: Punnett Square for second generation White Eyed Male and Wild Type Female Cross Parent Generation: XWt XW x XWt Y Wild Type XWt Wild Type XWt Wild Type Y XWt XWt XWt Y Wild Type XW XW XWt XW Y .
but also successful at putting the flies to sleep but not killing them. The tubing was then placed into the white eyed tube. 4. The tube was labeled using a piece of paper to indicate the type of cross contained within the tube. and one fourth white eyed male. a plastic tube was half filled with water. The CO2 from the alka seltzer and the water successfully put the flies to sleep. This method of anesthesia was used because it is inexpensive. The stock tube for white eyed flies was anesthetized using a chryalizer. Stock tubes for both wild type and white eyed flies were allowed to reproduce to provide an ample amount of flies for experimentation. one scoop of water. 2. one fourth wild type female. .X LINKED TRAITS OF WHITE EYED MALE AND WILD TYPE FEMALE CROSS 6 Figure two shows that the expected outcome for the second generation was half wild type male. To do this. and three grains of yeast into the tube. Materials Wild type fruit fly stock tubes Media Water Cryaliser Tape White eyed fruit fly stock tubes Yeast Foam Toppers Alka Seltzer Tube Labels Plastic tubes for f1 and f2 generations Procedure 1. and a cork with tubing was placed over the tube. 3. Half of an alka seltzer tablet was then dropped into the tube. The tube for F1 generation was made by putting one scoop of yeast.
After 6 hours. and then the flies were disposed of. a fly cannot become fertile for 8 hours. The f1 generation was anesthetized using the same procedure described in step 4. These five male white eyed flies were placed into the f1 stock tube to become the males in the parental generation of the f1 tube. Of the flies in the petri dish. 5 males and 5 females were taken and placed into the f2 tube. 10. Approximately 6 hours after the tube was emptied. 6. they were poured into a petri dish and five males were found. the f2 stock tube was created using the methods described in steps 2-3. and the flies were placed into a petri dish. 8. . After 100 flies had been created. After hatching from an egg. 9. The flies were retrieved within 6 hours to ensure their virginity. 12. Next. the stock for the wild type fruit flies was emptied.X LINKED TRAITS OF WHITE EYED MALE AND WILD TYPE FEMALE CROSS 7 5. 3 of these females were retrieved and placed into the f1 stock tube to become the white eyed females for the f1 generation. These offspring from the f1 would be the parent generation for the f2 tube. the newly hatched flies were anesthetized using the same method as described in step 4. leaving only the larvae left. After the flies were asleep. the dead flies were taken out of the freezer. and thus would be virgins. Retrieving them within 6 hours ensures they could not have mated yet. The flies were then given time to reproduce until 100 flies had been created. 13. as this would alter the accuracy of the experiment. The remaining flies were placed in the freezer for approximately 6 hours to ensure their death. 11. This is important to ensure that no male wild type flies get into the f1 cross. and their eye color as well as sex were recorded in a data sheet. 7.
the dead flies were removed and counted. The f2 generation was given enough time to reproduce and produce 100 flies. 16. and virgin female wild type flies. If three types of flies were in the tube. This setup ensured that crossbreeding between different sexes and different mutations did not occur.X LINKED TRAITS OF WHITE EYED MALE AND WILD TYPE FEMALE CROSS 8 14. the sex of the parental generation makes a difference in the expected outcome for the f1 and f2 generations. After 6 hours. Setting up reciprocal crosses ensured that conclusive data could be attained. . The f1 tubes contained male white eyed flies. and therefore the sex of the parental generation is important and must be controlled. Their eye color as well as sex was recorded in a data sheet. The wild type flies had to be virgin to ensure that they did not give birth to a male wild type fly. Additionally. only one type of fly was placed in the tube besides wild type to ensure only one type of breeding was occurring. Because the white eye genetic mutation is a sex linked trait. and thus final results would not be conclusive. if both males and females were put into a tube. the offspring could not be predicted because the rate and manner of the mating between the three types of flies could not be controlled. This type of setup was necessary so that outside factors which could affect the accuracy of the results could be eliminated. This would alter the results as white eyed mutation is a sex linked trait. the type of mating was controlled. Reciprocal crosses were set up to ensure that the sex of the flies did in fact make a difference. By crossing only one type of mutation and one type of sex in the same tube. and ensured that the final results could not be affected by different types of mutations. Additionally. and then placed in a petri dish and into the freezer for 6 hours. After 100 flies were produced in the f2 generation. they were anesthetized as described in step 4. the offspring could not be traced to a specific parent. 15.
00 with 1 degree of freedom. .X LINKED TRAITS OF WHITE EYED MALE AND WILD TYPE FEMALE CROSS 9 The setup in this experiment eliminated these possible variables.00 0. Results F1 Cross: Parental generation: XWt XWt x XW Y Expected genotypic ratio: 50% XWt XW : 50% XWt Y Expected phenotypic ratio: 50% Wild Type Males : 50% Wild Type Females Observed phenotypic ratio: 50% Wild Type Males : 50% Wild Type Females Table 1: Recorded Fly Characteristics in f1 Generation Cross Generation Traits Sex Expected Observed (n = 100) White Eyed Male x Wild Type Female F1 White Eyed Wild Type Male Female Male Female 0 0 50% = 50 50% = 50 0 0 50 50 Table 2: Chi Square Calculations for f1 Data Fruit Fly Phenotype White Eyed Male White Eyed Female Wild Type Male Wild type Female Total Observed # Expected # Observed ± Expected 0 0 0 0 (Observed ± Expected) 2 0 0 0 0 (Observed # Expected #)2 Expected # 0.00 0.00 X2 = 0.00 0.00 0 0 50 50 100 0 0 50 50 Df = (n-1) = (2-1) = 1 The Chi Squared value equals 0. and thus ensured that only the hypothesis was being tested.
00 2. .966 27 0 37 52 100 29 0 29 58 Df = (n-1) = (3-1) = 2 The Chi Squared value equals 2.X LINKED TRAITS OF WHITE EYED MALE AND WILD TYPE FEMALE CROSS 10 F2 Cross: Parental generation: XWt XW x XWt Y Expected genotypic ratio: 25% XWt XWt : 25% XW XWt : 25% XWt Y : 25% XW Y Expected phenotypic ratio: 50% Wild Type Female : 25% Wild Type Male : 25% White Eyed Male Observed phenotypic ratio: 45% Wild Type Female: 32% Wild Type Male : 23% White Eyed male Table 3: Recorded Fly Characteristics in f2 Generation Cross Generation Traits Sex Expected Observed (n = 116) White Eyed Male x Wild Type Female F2 White Eyed Wild Type Male Female Male Female 25% = 29 0 25% = 29 50% = 58 27 0 37 52 Table 4: Chi Square Calculations for f2 Data Fruit Fly Phenotype White Eyed Male White Eyed Female Wild Type Male Wild type Female Total Observed # Expected # Observed ± Expected -2 0 8 -6 (Observed ± Expected) 2 4 0 64 36 (Observed # Expected #)2 Expected # 0.621 X2 = 2.966 with 2 degrees of freedom.207 0.138 0.
P = < 0. P = < 0.X LINKED TRAITS OF WHITE EYED MALE AND WILD TYPE FEMALE CROSS 11 . The f2 data was also tested for statistical significance using a Chi Square test (see table 4 and table 3).5).25).966 df = 2.Figure 3: Chi Squared Value Chart Conclusion The f1 data was tested for statistical significance using a Chi Square test (see table 2 and figure 3).00 df = 1. . The Chi Square test rejected the null hypothesis (X2 = 2. The Chi Square test rejected the null hypothesis (X2 = 0.
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