April Hallock SCIN 211

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Jan 9, 2024

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12/17/23, 7 : 06 PM Transgenic Fly Virtual Lab Lab Notebook Page 1 of 1 https://media.hhmi.org/biointeractive/vlabs/transgenic_fly/content/sect1_part6_quiz_results.html Part 6: Quiz answers 1. How is light produced by the transgenic flies? A. The same way that light is produced in fireflies. B. The transgenic flies have eaten a substance that produces light. C. Transcription of the transgene results in production of luciferase, which acts on luciferin. D. Turning on the room lights causes light to be produced by the flies. Correct! Your answer: C. C. CORRECT. Transcription of the transgene results in production of luciferase, which acts on luciferin. (Click here for more details.) 2. Maximal light output from the transgenic fly occurs when the promoter of which gene is activated? A. period B. luciferase C. mini-white D. timeless Correct! Your answer: A. A. CORRECT. In the virtual lab experiment, only the period promoter is adjacent to the luciferase gene. When the period promoter is activated, luciferase gene transcription is turned on, producing luciferase mRNA. Luciferase mRNA is translated into luciferase protein, which acts on luciferin (in the fly's diet) to produce light. View lab status and quiz summaries

12/17/23, 6 : 53 PM Transgenic Fly Virtual Lab Lab Notebook Page 1 of 2 https://media.hhmi.org/biointeractive/vlabs/transgenic_fly/content/sect1_part4_quiz_results.html Part 4: Quiz answers 1. After completing a transgenic injection experiment, only white-eyed progeny result. Which of the following situations could explain the result? A. You did not inject enough embryos. B. You accidentally broke the glass needle and had to replace it with a new one in the micromanipulator. In the meantime, the embryos continued to develop, and all were more than 60 minutes old when you began the injections. C. All your injections missed the germ cells in the posterior region of the embryos. D. All the above. Correct! Your answer: D. D. CORRECT. All the above. A. INCORRECT. Even if you inject a few embryos correctly, all these embryos can die. A second possibility is that surviving embryos may have been injected with construct DNA but that DNA may not have integrated into the embryos' DNA. Both possibilities could result in only white-eyed progeny. Therefore, it is important to correctly inject as many embryos as possible. B. INCORRECT. All reagents and supplies need to be prepared ahead of time so the embryos can be injected promptly when they are less than 30 minutes old. C. INCORRECT. To obtain red-eyed flies, the DNA must be injected into the cells that will become germ cells, located at the posterior end of the embryo. 2. All the injected embryos that have grown to adulthood have white eyes. They are mated with wild-type flies that also have white eyes. How is it possible that some progeny will have red-eyes? A. Red-eyed flies contaminate the experiment. B. Progeny with red-eyes are the result of matings in which the germ cell from one parent contains the transgene. C. A mutation occurs that produces red eyes. D. Progeny with red eyes occur in flies raised in constant darkness. Correct! Your answer: B. B. CORRECT. Progeny with red-eyes are the result of matings in which the germ cell from one parent contains the transgene. A. INCORRECT. These experiments are conducted under carefully controlled conditions such that no contamination is possible. Print

12/17/23, 6 : 50 PM Transgenic Fly Virtual Lab Lab Notebook Page 1 of 2 https://media.hhmi.org/biointeractive/vlabs/transgenic_fly/content/sect1_part3_quiz_results.html Part 3: Quiz answers 1. Injected fly embryos fail to survive for all of the reasons below except A. fluid loss due to wound at injection site B. injection of too much fluid, causing embryo to explode C. embryo is too young to tolerate injection D. embryo becomes too dry Correct! Your answer: C. C. CORRECT. Younger embryos actually tolerate the procedure better than do older embryos. A, B, and D. INCORRECT. Excessive fluid loss, injecting too much DNA solution such that the cell explodes, and embryos drying out all may result in the death of the embryo. 2. When injecting flies with the construct DNA, which of the following steps will help increase the chances that transgenic progeny are produced? A. Inject a large volume of DNA solution. B. Inject many embryos. C. Inject into the outermost embryonic membrane. D. Destroy all older embryos. Correct! Your answer: B. A. INCORRECT. Injecting too large a volume of DNA solution will cause the embryo to explode, as illustrated in the interactive window if the mouse button is clicked more than once. B. CORRECT. Injecting many embryos will increase the likelihood that transgenic progeny are produced. However, the injection must occur in a specific time window during embryonic development, limiting the number of embryos that can be injected at once. C. INCORRECT. The needle must penetrate all embryonic membranes and enter the interior region of the embryo where cells will become germ cells. D. INCORRECT. The researcher may destroy older embryos, because these embryos are too old to incorporate the injected DNA into their cells. This procedure limits the number of adults with non transgenic germ cells, reducing the number of white-eyed non transgenics to sort through in later steps. However, this procedure does nothing to improve the likelihood of obtaining transgenic progeny.

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12/17/23, 6 : 45 PM Transgenic Fly Virtual Lab Lab Notebook Page 1 of 2 https://media.hhmi.org/biointeractive/vlabs/transgenic_fly/content/sect1_part1_quiz_results.html Part 1: Quiz answers 1. In this experiment, the reporter gene A. tells the experimenter exactly how much of the native protein was produced B. changes the color of the flies' eyes C. must be from the same type of organism as the organism into which it is being transferred D. is designed to be easily detected and reflect the activity of the native gene Correct! Your answer: D. A. INCORRECT. Levels of native protein (in this case, endogenous period protein) are not reported by the reporter gene. B. INCORRECT. The reporter gene in this experiment is luciferase , which produces light in amounts proportional to the transcriptional activity of the per promoter. The marker gene mini-white is responsible for the red eye color in those flies that have incorporated the DNA construct. C. INCORRECT. Reporter genes can originate from many sources. For example, the firefly luciferase gene has been inserted into plants, and the gene for the jellyfish green fluorescent protein has been inserted into mice. D. CORRECT. Reporter genes are designed to be easily measured and quantified, either by eye or by machines. 2. After successfully incorporating into the genome of a fly, the construct DNA will A. change the functions of the native per gene B. cause the eyes to be white C. reflect transcriptional activity without interfering with functions D. alter the biological clock Correct! Your answer: C. C. CORRECT. The transgenic fly is expected to have normal per gene activity and a normal biological clock. Two differences occur as a result of incorporation: the transgenic fly has red eyes and per transcriptional activity is reported as light emissions. A, B, and D. INCORRECT. The transgenic fly is expected to have normal per gene activity and a normal biological clock. Two differences occur as a result of incorporation: the transgenic fly has red eyes and per transcriptional activity is reported as light emissions.

12/17/23, 6 : 47 PM Transgenic Fly Virtual Lab Lab Notebook Page 1 of 1 https://media.hhmi.org/biointeractive/vlabs/transgenic_fly/content/sect1_part2_quiz_results.html Part 2: Quiz answers 1. Why is it important to monitor the age of the fly embryos? A. Cell membranes form around each nucleus in the multi-nucleated embryonic cell mass at approximately 30—60 minutes. B. Germ cells can incorporate DNA before differentiation, which begins when the embryo is approximately 30—60 minutes old. C. Embryos older than 30—60 minutes will not incorporate the DNA and will dilute the number of successfully injected flies. D. All of the above. Correct! Your answer: D. D. CORRECT. 2. An experimentally useful per-luc transgenic fly is produced A. when the injected fly embryo becomes an adult B. in all the offspring of an injected embryo that has grown to adulthood C. in only a few of the offspring of an injected embryo containing germ cells that have incorporated the injected DNA D. None of the above. Correct! Your answer: C. A, B, and D. INCORRECT. The transgene is not necessarily incorporated into all of the germ cells. It is possible that no germ cells, or only a few, may contain the transgene. Therefore, only a few of the progeny of the injected embryo may be per-luc transgenic flies. In this experiment, the presence of the transgene is confirmed when red-eyed flies are found. (Note that it is possible for the injected fly to contain the transgene in other cells as well; however, the presence of the transgene in other cells is irrelevant because only the transgene found in the sperm or eggs will be passed on to it's progeny.) View lab status and quiz summaries Print

12/17/23, 7 : 08 PM Experiment 1: Graph Analysis Page 1 of 1 https://media.hhmi.org/biointeractive/vlabs/transgenic_fly/content/sect2_exp1_analysis.html Experiment 1: Graph Analysis Mouse over (or tap) the peaks and troughs of the graph to display the corresponding times. Record the peak and trough times in the table below the chart. When you are finished recording, print this window and then click the Take the Quiz link at the bottom of this page. Note: you may leave this window open to refer to your analysis when taking the quiz. If you close this window, you will lose your data. Printing this window is recommended. Normal 12-hours-light/12-hours-dark schedule: Daytime is 6:00 a.m. — 6:00 p.m. Day Time of Peak (Max.) Time of Trough (Min.) 1 2 3 4 5 Dark-dark schedule: Lights off Day Time of Peak (Max.) Time of Trough (Min.) 6 7 8 9 10 Print This Window then Take the Quiz

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