General Biology 2 Qtr 3 Week 1 PDF

Republic of the Philippines Department of Education Region VI- Western Visayas PAVIA NATIONAL HIGH SCHOOL SENIOR HIGH SCHOOL DEPARTMENT...

Republic of the Philippines Department of Education Region VI- Western Visayas PAVIA NATIONAL HIGH SCHOOL SENIOR HIGH SCHOOL DEPARTMENT GENERAL BIOLOGY I Name: __________________________Grade/Strand & Section: _______________ Date: _________ RECOMBINANT DNA (Quarter 3, Week 1) Learning Competencies with Code Outline the processes involved in genetic engineering (STEM_BIO11/12-IIIa-b-6) Discuss the applications of recombinant DNA (STEM_BIO11/12-IIIa-b-7) Background Information for the Learners Definition of Terms Genetic Engineering  is the process of using recombinant DNA (rDNA) technology to alter the genetic makeup of an organism. It involves the direct manipulation of one or more genes. A gene from another species is added to an organism's genome to give it a desired phenotype (National Human Genome Research Institute).  is also known as gene modification. It is the process of altering the DNA in an organism’s genome. It may mean changing one base pair ( A-T or C-G), deleting a whole region of DNA, introducing an additional copy of a gene, or extracting DNA from another organism’s genome and combining it with the DNA of that individual (yourgenome.org).  involves the use of molecular techniques to modify the traits of a target organism. The modification of traits may involve introduction of new traits into an organism, enhancement of a present trait by increasing the expression of the desired gene, and enhancement of a present trait by disrupting the inhibition of the desired gene’s expression (Commission on Higher Education, 2016). Plasmid  is a small, often circular DNA molecule found in bacteria and other cells. Plasmids are separate from the bacterial chromosome and replicate independently of it. They may be passed between different bacterial cells (National Human Genome Research Institute). Vector  is any vehicle, often a virus or a plasmid, used to ferry a desired DNA sequence into a host cell as part of a molecular cloning procedure. Depending on the purpose of the cloning procedure, the vector may assist in multiplying, isolating, or expressing the foreign DNA insert (National Human Genome Research Institute). Restriction enzyme  is an enzyme isolated from bacteria that cuts DNA molecules at specific sequences. The isolation of these enzymes was critical to the development of recombinant DNA (rDNA) technology and genetic engineering (National Human Genome Research Institute). Cloning  is the process of making identical copies of an organism, cell, or DNA sequence. Molecular cloning is a process by which scientists amplify a desired DNA sequence. The target sequence is isolated, inserted into another DNA molecule (known as a vector), and introduced into a suitable host cell. Then, each time the host cell divides, it replicates the foreign DNA sequence along with its own DNA. Cloning also can refer to asexual reproduction (National Human Genome Research Institute). Transgenic or Genetically Modified Organisms  means that one or more DNA sequences from another species have been introduced by artificial means. Animals usually are made transgenic by having a small sequence of foreign DNA injected into a fertilized egg or developing embryo. Transgenic plants can 1  be made by introducing foreign DNA into a variety of different tissues (National Human Genome Research Institute). Genome Editing  is a method that lets scientists change the DNA of many organisms, including plants, bacteria, and animals. Editing DNA can lead to changes in physical traits, like eye color, and disease risk (National Human Genome Research Institute).  is also called gene editing. It is a group of technologies that give scientists the ability to change an organism’s DNA. These technologies allow genetic material to be added, removed, or altered at particular locations in the genome. Several approaches to genome editing have been developed. A recent one is known as CRISPR-Cas9, which is short for clustered regularly interspaced short palindromic repeats and CRISPR- associated protein 9. The CRISPR-Cas9 system has generated a lot of excitement in the scientific community because it is faster, cheaper, more accurate, and more efficient than other existing genome editing methods (Medline Plus). Gene therapy  is an experimental technique for treating disease by altering the patient's genetic material. Most often, gene therapy works by introducing a healthy copy of a defective gene into the patient's cells (National Human Genome Research Institute). Processes Involved in Genetic Engineering A general outline of recombinant DNA may be given as follows: I. cutting or cleavage of DNA by restriction enzymes (REs) II. selection of an appropriate vector or vehicle which would propagate the recombinant DNA ( eg. circular plasmid in bacteria with a foreign gene of interest) III. ligation (join together) of the gene of interest (eg. from animal) with the vector ( cut bacterial plasmid) IV. transfer of the recombinant plasmid into a host cell (that would carry out replication to make huge copies of the recombined plasmid) V. selection process to screen which cells actually contain the gene of interest VI. sequencing of the gene to find out the primary structure of the protein (Commission on Higher Education, 2016). Ways in which plasmids may be introduced into host organisms 1. Biolistics A “gene gun” is used to fire DNA-coated pellets on plant tissues. Cells that survive the bombardment are able to take up the expression of plasmid coated pellets and acquire the ability to express the designed protein. 2. Plasmid insertion by Heat Shock Treatment Heat Shock Treatment is a process used to transfer plasmid DNA into bacteria. The target cells are pre-treated before the procedure to increase the pore sizes of their plasma membranes. This pretreatment (usually with CaCl2) is said to make the cells “competent” for accepting the plasmid DNA. After the cells are made competent, they are incubated with the desired plasmid at about 4°C for about 30min. The plasmids concentrate near the cells during this time. Afterwards, a “Heat Shock” is done on the plasmid-cell solution by incubating it at 42°C for 1 minute then back to 4°C for 2 minutes. The rapid rise and drop of temperature is believed to increase and decrease the pore sizes in the membrane. The plasmid DNA near the membrane surface are taken into the cells by this process. The cells that took up the plasmids acquire new traits and are said to be “transformed”. 3. Electroporation It follows a similar methodology as Heat Shock Treatment, but the expansion of the membrane pores is done through an electric “shock”. This method is commonly used for insertion of genes into mammalian cells. Methods to Screen Recombinant Cells 1. Selection of plasmid DNA containing cells A selection marker within the inserted plasmid DNA sequence allows the selection of “transformants”. Usually, an antibiotic resistance gene (e.g. AMP ampicillin resistance gene) is included in the plasmid DNA. This allows only “transformed” cells to survive in the presence of the antibiotic (e.g. ampicillin). Plating the plasmid-cell solution 2  on antibiotic-containing media will select for these “transformants” and only allow plasmid-containing cells to grow and propagate into colonies. 2. Selection of transformed cells with the desired gene Certain inserted genes within the plasmids provide visible proof of their presence. These include the antibiotic resistance genes that allow for the selection of the transformed cells within the solution. Some inserted genes also produce colored (e.g. chromogenic proteins) or fluorescent products) or (e.g. GFP) that label the colonies/cells with the inserted gene. 3. PCR detection of plasmid DNA Alternatively, the presence of the desired gene in the inserted plasmids may be confirmed using PCR amplification. PCR reactions specific for the desired gene may be done using DNA from cells. Amplification of the expected product would confirm the presence of the gene within the samples. PCR reactions specific for plasmid sequences will also confirm/identify the type of plasmid used for the transformation. The Genetic Engineering Process (Example: Production of Human Insulin) 1. A small piece of circular DNA called a plasmid is extracted from the bacteria or yeast cell. 2. A small section is then cut out of the circular plasmid by restriction enzymes (molecular scissors). 3. The gene for human insulin is inserted into the gap in the plasmid. This plasmid is now genetically modified. 4. The genetically modified plasmid is introduced into a new bacteria or yeast cell. 5. This cell then divides rapidly and starts making insulin. 6. To create large amounts of the cells, the genetically modified bacteria or yeast are grown in large fermentation vessels that contain all the nutrients they need. The more the cells divide, the more insulin is produced. 7. When fermentation is complete, the mixture is filtered to release the insulin. 8. The insulin is then purified and packaged into bottles and insulin pens for distribution to patients with diabetes. (yourgenome.org) 3  Applications of Recombinant DNA Technology 1) Food and agriculture  Manufacture of novel enzymes which are suitable in food industries which includes lipases and amylases  Development of microbial strains  Produce enzymes for production of proteases  Strains of fungi had been modified in order to have reduced ability to produce toxic materials  Inhibits food spoilage  Recombinant proteins are used as enzymes in industries  Proteins in milk play a role in nutrition  Polymeric proteins are used in industries and medical field  Hepatitis B Vaccine production in plants  Oral vaccination with edible plants  Plants are used to produce several therapeutic protein products, such as casein and lysozyme for improving health of child and polymers of proteins for tissue replacement and surgery.  Tobacco plants can be engineered genetically to produce human collagen.  Chloroplast genome sequences are important in plant evolution and phylogeny.  Lycopene β-cyclase genes introduction into the plastid genome of tomato enhances the lycopene conversion into provitamin A.  Resistance to fungal and bacterial infections  Potato, beans, eggplant, sugar beet, squash, and many other plants are being developed with desirable characters, for example, tolerance of the herbicide glyphosate, resistance to insects, drought resistance, disease and salt tolerance. Nitrogen utilization, ripening, and nutritional versatility like characters have also been enhanced. 2) Health and Diseases  Gene Therapy  Treatment for adenosine deaminase-deficiency ( ADA-SCID)  Many different cancers including lung, gynecological, skin, urological, neurological, and gastrointestinal tumors, as well as hematological malignancies and pediatric tumors, have been targeted through gene therapy. Inserting tumor suppressor genes to immunotherapy, oncolytic virotherapy and gene directed enzyme prodrug therapy are different strategies that have been used to treat different types of cancers.  Will provide a new avenue for therapeutic angiogenesis, myocardial protection, regeneration and repair, prevention of restenosis following angioplasty, prevention of bypass graft failure, and risk-factor management  Viral gene therapy is one of the leading and important therapies for head and neck cancer.  Production of Antibodies and Their Derivatives  Transgenic tobacco plants can be used for the production of chimeric secretory IgA/G known as CaroRx. Oral pathogen responsible for decay of a tooth known as Streptococcus mutants, can be recognized by this antibody.  A monoclonal antibody called T84.66 can affectively function to recognize antigen carcinoembryonic, which is still considered an affectively characterized marker in cancers of epithelia  A full-length humanized IgG1 known as anti-HSV and anti-RSV, which can function as the recognizing agent for herpes simplex virus (HSV)-2- glycoprotein B, has been expressed in transgenic soybean and Chinese Hamster Ovary (CHO) cells.  Investigation of the Drug Metabolism  Recombinant DNA approaches have recently contributed its role through heterologous expression, where the enzyme's genetic information is expressed in vitro or in vivo, through the transfer of gene 4   Development of Vaccines and Recombinant Hormones  In vitro production of human follicle-stimulating hormone (FSH) is now possible through recombinant DNA technology.  r-FSH and Luteinizing Hormone (LH) recombination was made successful to enhance the ovulation and pregnancy.  Chinese Medicines  Chinese medicines play a crucial role in diagnostics and therapeutics  Medically Important Compound in Berries  Improvement in nutritional values of strawberries has been carried through rolC gene. This gene increases the sugar content and antioxidant activity. 3.Environment  Phytoremediation and Plant Resistance Development  used for the detection and absorption of contaminants in drinking water and other samples  Energy Applications  Cyanobacteria can be engineered to make them able to convert of CO 2 into reduced fuel compounds. (Khan et al., 2016) Directions / Instructions Read and study the different activities. Write your answer on a separate sheet of paper. Exercises / Activities Activity 1. What Promotes the Change? Tell whether the desirable traits had been a product of recombinant DNA technology or classical breeding. (1point each) Organism Enhanced Trait Modifying Technique Bt Eggplant Fruit- borer resistant Guapple Large-sized guava Golden Rice Vitamin A-enriched rice Papaya Delayed ripening Coconut Macapuno trait Tobacco Glow in the dark 5 Guide Questions (2 pts each) 1. What is classical breeding? 2. What is recombinant DNA technology? 3. What are the differences between classical breeding and recombinant DNA technology? Activity 2. The Making of a Bt Corn Sequence the following steps in the process of producing the Bt corn. Write number 1 on the first step and number 5 on the last step on the blank provided after each statement. (1 point each) In the laboratory, the specific DNA sequence which makes up the cry 1Ab _________ gene is found and copied. In a laboratory, a corn plant is transformed with this new modified cry 1Ab _________ gene which will provide the corn plant with insecticidal protection. All of the DNA from Bacillus thuringiensis, is isolated in a laboratory. _________ The corn plant which is able to be transformed does not contain all of _________ the optimal genes which a producer needs in the field. Therefore, plant breeders cross this corn plant (which contains the cry 1Ab gene) with their top performing lines to create a high yielding BT corn variety. The cry 1Ab soil bacterium version of the gene is modified slightly so _________ that it will work better once in a corn plant. Guide Questions (1 point each) 1. What is the gene of interest? 2. What is the source of the gene of interest? 3. What desirable trait is being expressed by the gene of interest? 4. What organism will receive the gene of interest? 5. What will be the result of this process? Activity 3a. Designing GMO (50 points) 1. Think of the organism with a trait that you want to improve. 2. Choose the organism from which you will get this gene that shows the desired trait. 3. Imagine what your organism would look like if the desired new trait is present. 4. Draw the resulting GMO you have designed. 5. Describe the characteristics of your GMO. 6. Give it a unique name. 7. Submit your work in our messenger group specifically created for GMO. Activity 3b. Making Decisions (30 points) Gene editing has the potential to alter traits such as intelligence and beauty in humans. If given a choice, will you submit yourself for gene editing for that purpose? Why or why not? Activity 4. Performance Task (100 points) Make a research paper, case study or poster on genetic diseases. Instructions and rubric will be posted in the google classroom. 6 Reflections Complete the statements below.( 2 points each) I understand ________________________________________________________ ________________________________________________________ I don’t understand _______________________________________________________ _______________________________________________________ I need more information about ________________________________________________________ ________________________________________________________ Answer Key Activity 1 Activity 2 1.Recombinant DNA Technology 2, 4,1,5,3 2.Classical Breeding Guide Questions 3. Recombinant DNA Technology 1.cry 1Ab 4. Recombinant DNA Technology 5. Classical Breeding 2. Bacillus thuringiensis 6. Recombinant DNA Technology 3. insect resistant/ insecticidal protection Guide Questions 4. corn 1-3. Answers may vary 5. insect-resistant corn plant Activities 3a,3b and 4 Answers may vary References Commission on Higher Education (2016). Teaching Guide for Senior High School General Biology 2 Khan,S.et al., ( 2016). Role of recombinant DNA technology to improve life. International Journal of Genomics doi.org/10.1155/2016/2405954, Retrieved from https://www.hindawi.com/journals/ijg/2016/2405954/ https://www.genome.gov/genetics-glossary/Genetic-Engineering https://www.britannica.com/science/genetic-engineering https://www.yourgenome.org/facts/what-is-genetic-engineering https://medlineplus.gov/genetics/understanding/genomicresearch/genomeediting/ https://www.ncbi.nlm.nih.gov/books/NBK21756/ https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5178364/ https://cropwatch.unl.edu/biotechnology/makinggmo Note. Please practice personal hygiene at all times. Prepared by: SIENA G. CAJILIG, PhD Master Teacher II 7