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8 Biotechnology Quarter 3 – Module 2: Genetic Manipulation Biotechnology– Grade 8 Alternative Delivery Mode Quarter 3 – Module 2: Genetic Manipulation First Edition, 2021 Republic Act 8293, section 176 states that: No copyright shall subsist in any work of the Government of the Philippines. However, prior approval of the government agency or office wherein the work is created shall be necessary for exploitation of such work for a profit. Such agency or office may, among other things, impose as a condition the payment of royalties. Borrowed materials (i.e., songs, stories, poems, pictures, photos, brand names, trademarks, etc.) included in this module are owned by their respective copyright holders. Every effort has been exerted to locate and seek permission to use these materials from their respective copyright owners. The publisher and authors do not represent nor claim ownership over them. Regional Director : OIC Asst. Regional Director : May B. Eclar PhD, CESO V Rhoda T. Razon EdD, CESO V Development Team of the Module Writers: Marifar Santos, Lemuel Licup, Larissa Manalili Editors: Sherilyne L. Reyes, Jennifer Praza, Edgardo D. Cortez, Jenny S. Tongol, Edythe Hipolito Reviewers: Gemima A. Estrabillo, Emily F. Sarmiento, Hermes Vargas, Adrian Tamayo, Krislene Ida N. Mercado, Noel S. Reganit Mercedes Bactol, Billy Ray B. Manuel, Marvin R. Leano, Gemmarie G. Rivas Illustrator: Arnold Arceo Layout Artist: Maricon H. Rivera, Noel S. Reganit Management Team: May B. Eclar PhD, CESO V Rhoda T. Razon EdD, CESO V Ma. Irelyn P. Tamayo PhD, CESE Fernandina P. Otchengco, PhD, CESE Librada M. Rubio PhD Ma. Editha R. Caparas EdD Rochella C. David Emily F. Sarmiento PhD Gemima A. Estrabillo EdD Printed in the Philippines by Department of Education –Region III – Schools Division of Angeles City Office Address: Jesus St., Pulungbulu, Angeles City Telefax: (045) 322-5722; 322-4702; 888-0582; 887-6099 E-mail Address: [email protected] 8 Biotechnology Quarter 3 – Module 2: Genetic Manipulation Introductory Message This Self-Learning Module (SLM) is prepared so that you, our dear learners, can continue your studies and learn while at home. Activities, questions, directions, exercises, and discussions are carefully stated for you to understand each lesson. Each SLM is composed of different parts. Each part shall guide you step-bystep as you discover and understand the lesson prepared for you. Pre-tests are provided to measure your prior knowledge on lessons in each SLM. This will tell you if you need to proceed on completing this module or if you need to ask your facilitator or your teacher’s assistance for better understanding of the lesson. At the end of each module, you need to answer the post-test to self-check your learning. Answer keys are provided for each activity and test. We trust that you will be honest in using these. In addition to the material in the main text, Notes to the Teacher are also provided to our facilitators and parents for strategies and reminders on how they can best help you on your home-based learning. Please use this module with care. Do not put unnecessary marks on any part of this SLM. Use a separate sheet of paper in answering the exercises and tests. And read the instructions carefully before performing each task. If you have any questions in using this SLM or any difficulty in answering the tasks in this module, do not hesitate to consult your teacher or facilitator. Thank you. What I Need to Know This module was designed and written with you in mind. It is here to help you master the nature of Biotechnology. The scope of this module permits it to be used in many different learning situations. The language used recognizes the diverse vocabulary level of students. The lessons are arranged to follow the standard sequence of the course. But the order in which you read them can be changed to correspond with the textbook you are now using. The module is summated into one lesson, namely: • Lesson 1: Genetic Manipulation After going through this module, you are expected to discuss how genetic materials are manipulated; 1 What I Know Directions: Read each question carefully. Choose the letter of the correct answer. 1. This refers to the process of manipulating genes for practical purposes. a.Genetic engineering b. Karyotyping c. Ligase d. Plasmid 2. The circular piece of DNA found in bacteria and contains genes. a. Cell b. Ligase c. Plasmid d. Vector 3. The process by which many copies of a specific gene are made each time the host cell reproduces. a. Fingerprinting b. PCR c. Gene Cloning d. Recombinant DNA 4. It is the shape of the DNA. a. Helical c. Parallel b. horizontal d. Vertical 5. He discovered the Polymerase Chain Reaction. a. Hamilton Smith b. Hindll c. Kary Mullis d. Werner Abner 6. These are enzymes used to cut DNA at specific locations based on the nucleotide sequence. a. Gel electrophoresis b. Ligase c. Restriction enzymes d. PCR 7. It is a genetic engineering tool used to multiply the DNA exponentially for each of the 25 to 75 cycles. a. Gel electrophoresis b. Ligase c. Restriction enzymes d. PCR 8. It is a branch of biotechnology deals with the study and investigation of genomic information from trace evidence found at crime scenes. a. Agricultural biotechnology b. Forensic biotechnology c. Industrial biotechnology d. Medical biotechnology 9. This was the name given to the first mouse used to study cancer. a. Mighty mouse b. Mousegenic c. Oncomouse d. Supermouse 10. This is the process where mice are used to study gene function by purposely turning off specific genes. a. Apoptosis b. Cell knockout c. Gene knockout d. Genetosis 2 Lesson 1 Genetic Manipulation What’s In In our previous module, we learned about the different tools used in genetic engineering and how these tools became helpful for human in terms of crop production, detection of viruses, forensics and development of vaccines. For example, the reverse transcription polymerase chain reaction (rRt-PRC) test which is used to detect if a person is infected with a common strain of virus. In addition, the pulse field gel electrophoresis is a technique that distinguishes samples of genetic material applied in DNA fingerprinting. ACTIVITY A Direction: Match each tool used in genetic engineering to their corresponding description by writing on the blank spaces the letter of the correct answer. Genetic Tools Polymerases Gel Electrophoresis Eukaryotic Host Polymerase Chain Reaction Molecular Scissor Answer Description a. It multiplies the DNA exponentially for each of the 25 to 75 cycles. b. It is the enzymes that cut DNA at specific locations based on the nucleotide sequence. c. It monitors the changes protein content in body fluids. of d. The utilization of multi cell organisms to produce human proteins since these hosts with complex structures are more suitable to synthesize complex proteins. e. The groups of enzymes that catalyze the synthesis of nucleic acid molecules. 3 What’s New Directions: Study the flowchart below. Complete it by choosing inside the box the missing steps and benefits of DNA manipulation. Other responses were given as your clues. A- Removal of genes D- To produce hydrocarbons, fuels, plastics, drugs B- Mutation of existing genes E- To produce disease-and insect resistant crops, edible vaccines, larger crops C- Insert new genes G- To track protein production, for disease detection to produce larger animals as food source DNA Manipulation (Steps) 3. 2. Replace insert new genes 1. 4. Creates genetically modified organism GM Plants GM Bacteria GM Animals Benefits 5. 7. 6. 4 What is It Let’s talk about what is DNA, or deoxyribonucleic acid, is the hereditary material in humans and almost all other organisms. So now let’s learn what is genetic manipulation, it is the process of inducing changes in gene expression and expression of novel genes and has proven to be an indispensable tool in genetic research. According to Bhagvanji, Gohil Sanjay (2018) one of the applications of genetic manipulation is Gene cloning, defined as a mechanism by which each time the host cell reproduces; several copies of a particular gene are produced wherein it is possible to clone whole species. Gene Cloning is the insertion of a fragment of DNA carrying a gene into a cloning vector and subsequent propagation of recombinant DNA molecules into many copies is known as gene cloning. It involves using bacteria to make multiple copies of a gene, foreign DNA is inserted into a plasmid, and the recombinant plasmid is inserted into a bacterial cell, reproduction in the bacterial cell results in cloning of the plasmid including the foreign DNA and this results in the production of multiple copies of a single gene. Nuclear cloning or Nuclear transfer- the introduction of the nucleus from a cell into an enucleated egg cell (an egg cell that has had its own nucleus removed). This can be accomplished through fusion of the cell to the egg or through the direct removal of the nucleus from the cell and the subsequent transplantation of that nucleus into the enucleated egg cell. The donor nucleus used for nuclear transfer may come from an undifferentiated embryonic cell or from a differentiated adult cell (somatic cell); in the latter case, the technique is called somatic cell nuclear transfer. The concept of nuclear transfer was first conceived in 1928 by German embryologist 5 Hans Spemann, who initially experimented with transferring salamander embryonic cell nuclei into egg cells (Rogers, Kara). Transgenic organism- Modern genetic technology can be used to modify the genomes of living organisms. This process is also known as “genetic engineering.” Genes of one species can be modified, or genes can be transplanted from one species to another. Genetic engineering is made possible by recombinant DNA technology. Organisms that have altered genomes are known as transgenic. Most transgenic organisms are generated in the laboratory for research purposes. For example, “knock-out” mice are transgenic mice that have a particular gene of interest disabled. By studying the effects of the missing gene, researchers can better understand the normal function of the gene (Transgenic Organisms. 2019. Genetics Generation). Transgenic organisms have also been developed for commercial purposes. Perhaps the most famous examples are food crops like soy and corn that have been genetically modified for pest and herbicide resistance. These crops are widely known as “GMOs” (genetically modified organisms). Here are few other examples of transgenic organisms with commercial value: Golden rice: modified rice that produces beta-carotene, the precursor to vitamin A. Vitamin A deficiency is a public health problem for millions of people around the world, particularly in Africa and Southeast Asia. Golden rice is still waiting regulatory approval. Goats that produce important proteins in their milk: goats modified to produce FDA-approved human antithrombin (ATryn), which is used to treat a rare blood clotting disorder in humans. Goats have also been genetically modified to produce spider silk, one of the strongest materials known to man, in their milk. Proposed uses for this recombinant spider silk range from artificial tendons to bulletproof vest. Copyright 1999-2009 Access Excellence @ The National Health Museum. Alright reserved. 6 The process of cloning a gene in a bacterial plasmid can be divided into five steps: 1. Isolation of vector and gene-source DNA. The source DNA may come from human tissue cells. The source of the plasmid is typically E. coli. This plasmid carries useful genes, such as ampR, conferring resistance to the antibiotic ampicillin. 2. Insertion of DNA into the vector. By digesting both the plasmid andhumanDNA with the same restriction enzyme we can create thousands of human DNA fragments, one fragment with the gene that we want, and with compatible sticky ends on bacterial plasmids. After mixing, the human fragments and cut plasmids form complementary pairs that are then joined by DNA ligase. This creates a mixture of recombinant DNA molecules. 3. Introduction of the cloning vector into cells. Bacterial cells take up the recombinant plasmids by the transformation. This creates a diverse pool of bacteria, some bacteria that have taken up the desired recombinant plasmid DNA, other bacteria that have taken up other DNA, both recombinant and nonrecombinant. 4. Cloning of cells (and foreign genes). We can plate out the transformed bacteria on a solid nutrient medium containing ampicillin. Only bacteria that have the ampicillin-resistance plasmid will grow. 5. Identifying cell clones with the right gene. Inthe final step, we willsortthrough the thousands of bacterial colonies with foreign DNA to find those containing our gene of interest Transgenic animals are used to study diseases and gene functions. Transgenic mice were used to study development and disease; the first mouse used was called an oncomouse used to study cancer. Other mice are used to study diabetes, brain function, and development and sex determination. Gene knockout mice used to study gene function – by purposely “turning off” specific genes. The knockout mouse does not have a functional gene for a protein called leptin, which helps to control food intake. Researchers are using this type of mouse to study obesity. 7 What’s More Activity 1 Dolly: The cloned sheep Direction: Read the short article about Dolly the sheep and answer the following guide questions. Dolly was part of a series of experiments at The Roslin Institute that was trying to develop a better method for producing genetically modified livestock. If successful, this would mean fewer animals would need to be used in future experiments. Scientists at Roslin also wanted to learn more about how cells change during development and whether a specialized cell, such as a skin or brain cell, could be used to make a whole new animal. These experiments were carried out at The Roslin Institute by a team led by Professor Sir Ian Wilmut. Because of the nature of the research, the team was made up of many different people, including scientists, embryologists, surgeons, vets and farm staff. Dolly was cloned from a cell taken from the mammary gland of a six-year-old Finn Dorset sheep and an egg cell taken from a Scottish Blackface sheep. She was born to her Scottish Blackface surrogate mother on 5th July 1996. Dolly’s white face was one of the first signs that she was a clone because if she was genetically related to her surrogate mother, she would have had a black face. In 2001, Dolly was diagnosed with arthritis after farm staff noticed her walking stiffly. This was successfully treated with anti-inflammatory medication, although the cause of the arthritis was never discovered. Dolly continued to have a normal quality of life until February 2003, when she developed a cough. A CT scan showed tumors growing in her lungs and the decision was made to euthanize Dolly rather than risk her suffering. Dolly was put to sleep on 14th February 2003, at the age of six. Guide questions: 1. Who is Dolly the Sheep and how was she created? 2. Who made Dolly the Sheep? 3. What was the first sign that showed Dolly was cloned? Assessment1. 1. Why do you think producing Dolly is an important breakthrough in Science and Technology? 2. If you were chosen to be a part of the team who created Dolly, what else would you consider so Dolly would be healthy as possible? 8 Activity 2. Directions: Complete the sentences in the diagram below to show the manipulation of genetic material. Photo source: April 30, 2018|Author: Anonymous| Category: Science, Biology, Biochemistry, Genetics Assessment 2. 1. Based on the diagram in activity 2, discuss the manipulation of genetic material on how they cloned a lamb. Activity 3. Gene Cloning versus Nuclear Cloning Direction: Copy the Venn Diagram below and write the similarities and differences between Gene cloning and Nuclear cloning. NUCLEAR CLONING GENE CLONING Differences Similarities Differences Assessment 3 1. What are the two types of Cloning? 2. What is Gene Cloning? How about Nuclear Cloning? 3. What type of cloning did scientists use to create Dolly? Explain your answer? 9 Activity 4: Directions: Read each description carefully. Match them with the terms used in DNA manipulation. Write the letter of the correct answer before each number. Descriptions Terms A. Transgenic Organism 1. It is the process of inducing changes in gene expression and expression of novel genes. B. Nuclear cloning 2. It is defined as a mechanism by which each time the host cell reproduces. C. Genetic manipulation D. Gene cloning 3. It is a modern technology which is used to modify the genomes of living organisms. E. Gene knockout 4. It is the purposely “turning off” the specific genes of a mouse. 5. It is the introduction of the nucleus from a cell into an enucleated egg cell. Assessment 4. 1. What is oncomouse used for? 2. Why do we create transgenic animals? 3. How does cloning benefit the environment? Activity 5: MYSTERY WORDS Directions: Arrange the jumbled letters to form correct words. Then, copy the letters in the numbered boxes and write them below to unlock the mystery words. 1 2 3 4 5 6 7 8 9 10 11 1. SILIONATO 2 2. COERTNISRTI 7 3. NROOSARFAITT 6 3 4. PIADLSM 1 10 10 12 13 14 15 5. MERTBCAONNI 4 6. SORSIPEEXN 8 7. GERTAT GEEN 11 8. NEZYSME 13 9. BEACIART 12 10. RIVUS 9 Assessment 5. 1. Based on the puzzle above, how are the terms involved in the DNA manipulation process? Discuss your answers. Activity 6. Letter clues Direction: Decode the terms related to DNA manipulation by corresponding letter of the given number clues. writing the A B C D E F G H I J K L M (1) (2) (3) (4) (5) (6) (7) (8) (9) (10) (11) (12) (13) N O P Q R S T U V W X Y Z (14) (15) (16) (17) (18) (19) (20) (21) (22) (23) (24) (25) (26) 1. It is the process by which many copies of a specific gene are made each time the host cell reproduces. 7-5-14-5-3-12-15-9-14-7 2. What do we call to the organisms created by genetic engineering? 7 – 13 – 15 3. It is the introduction of the nucleus from a cell into an enucleated egg cell. 14-213-12-5-1-18 4. It contains foreign DNA that has been introduced using biotechnology. 20-1- 1419-7-5-14-9-3 5. The transgenic animals that are used to study cancer. 15-14-3-15-13-15-21-19-5 11 Assessment 6. 1. Based on your previous activity, how can we differentiate genetically modified organisms and transgenic organisms? And how are transgenic organisms created? What I Have Learned Let’s sum up what you have learned. Complete the sentences below. It is the hereditary material in humans and almost all other organisms 1 . While 2. is the basic physical and functional unit of heredity and made up of DNA. 3. is the process of producing individuals with identical or virtually identical DNA, either naturally or artificially. There are 2 types of cloning the 4 and 5 . Organisms that have altered genomes are known as 6. which generated in the laboratory for research purposes. For example, 7 are transgenic mice that have a particular gene of interest disabled and used to study cancer. Another example is 8. the first mammal cloned from an adult somatic cell, using the process of nuclear. While 9. is a modified rice that produces 10. the precursor to vitamin A. 12 What I Can Do Find an article about the Human Genome Project. Write your insights based on the article that you have read. Use the rubric below to guide you in writing your content. CONTENT ORGANIZATION STYLE 5pts. The ideas are substantial, specific, and/or illustrative content demonstrating strong development and sophisticated ideas. Sophisticated arrangement of content with evident and/or subtle transitions. Precise, illustrative use of a variety of words and sentence structures. 4pts. Sufficiently developed content with adequate elaboration or explanation. Functional arrangement of content that sustains a logical order with some evidence of transitions. Use of simple or common words. 3pts. Limited content with inadequate elaboration or explanation. Confused or inconsistent arrangement of content with or without attempts at transition. Limited wordchoice and control of sentence structures. 2Pts. Superficial and/or minimal content. Minimal control of content arrangement. Minimal variety in word choice and minimal control of sentence structures. 13 Assessment Directions: Read each question carefully. Choose the letter of the correct answer 1. Which genetic engineering process contains foreign DNA that has been introduced using biotechnology? a. b. c. d. Gene cloning GMO Nuclear cloning Transgenic organism 2. Who developed the process of nuclear cloning? a. Hamilton Smith b. Hans Spemman c. Hindll d. Kary Mullis 3. When was the first nuclear cloning conceived? a. 1928 b. 1929 c. 1930 d. 1931 4. Which organism was modified to produce FDA-approved human antithrombin (ATryn). a. Cat b. Frog c. Goat d. Sheep 5. What vitamin is present in Golden rice? a. Vitamin A b. Vitamin B c. Vitamin C d. Vitamin D B. FACT OR BLUFF. Write Fact if the statement is True and write Bluff if it is False. 1. Dolly the sheep is still alive. 2. Transgenic organisms have also been developed for commercial purposes. 3. The donor nucleus used for nuclear transfer may come from an undifferentiated embryonic cell. 4. The new DNA can be inserted randomly, or targeted to a specific part of the genome. 5. DNA manipulation is the indirect manipulation of an organism's genes using biotechnology. 14 Additional Activity Direction: How does the creation of Spider goat be helpful? Discuss your answer. Photo source: by Lisa Zyga , Phys.org Rubric for Scoring CONTENT ORGANIZATION STYLE 5pts. The ideas are substantial, specific, and/or illustrative content demonstrating strong development and sophisticated ideas. Sophisticated arrangement of content with evident and/or subtle transitions. Precise, illustrative use of a variety of words and sentence structures. 4pts. Sufficiently developed content with adequate elaboration or explanation. Functional arrangement of content that sustains a logical order with some evidence of transitions. Use of simple or common words. 3pts. Limited content with inadequate elaboration or explanation. Confused or inconsistent arrangement of content with or without attempts at transition. Limited word choice and control of sentence structures. 2Pts. Superficial and/or minimal content. Minimal control of content arrangement. Minimal variety in word choice and minimal control of sentence structures. 15 References Anderson, Christine, and Lisa Bartee. n.d. “Manipulating Genetic Material.” Mt Hood Community College Biology 102. Open Oregon Educational Resources. Accessed January 16, 2021. Retrieved from https://openoregon.pressbooks.pub/mhccbiology102/chapter/manipulatin g-genetic-material/. Bhagvanji, Gohil Sanjay. 2018. “Gene Cloning Principles as a Technique.” Retrieved from SlideShare. https://www.slideshare.net/gohilsanjay3/gene-cloningprinciples-an-technique. Genetic Engineering. n.d. Retrieved from http://www.deercreekhs.org/UserFiles/Servers/Server_38468/File/Bush/H MH%209.4%20Notes.ppt Malaka Follow, Madhusudhana. 2019. “Gene Cloning Presentation.” SlideShare. Retrieved from https://www.slideshare.net/MadhusudhanaMalaka/genecloning-presentation. Rideout, William M., Kevin Eggan, and Rudolf Jaenisch. 2001. “Nuclear Cloning and Epigenetic Reprogramming of the Genome.” Science. American Association for the Advancement of Science. Retrieved from https://science.sciencemag.org/content/293/5532/1093. Rogers, Kara. n.d. “Nuclear Transfer.” Encyclopædia Britannica. Encyclopædia Britannica, inc. Accessed January 16, 2021. Retrieved from https://www.britannica.com/science/nuclear-transfer. The Life of Dolly. n.d. Dolly the Sheep. Accessed January 16, 2021. Retrieved from https://dolly.roslin.ed.ac.uk/facts/the-life-of-dolly/index.html. Transgenic Organisms. 2019. Genetics Generation. Retrieved from https://knowgenetics.org/transgenic-organisms/ 19 For inquiries or feedback, please write or call: Department of Education – Region III- Schools Division of Angeles City Jesus St., Pulungbulu, Angeles City, Pampanga, Philippines 2009 Telefax: (045) 322-5722; 322-4702; 888-0582; 887-6099 Email Address: [email protected]