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Chapter 5: DNA, Gene Expression, and Biotechnology What is the code and how is it harnessed? Lectures by Mark Manteuffel, St. Louis Community College; Clicker Questions by Kristen Curran, University of Wisconsin-Whitewater Learning Objectives Describe what DNA is and what it does. Explain the process of gene expression and the collaboration of nature and nurture. Explain the causes and effects of damage to the genetic code. Describe biotechnology and its implications for human health. Discuss biotechnology in agriculture. Discuss biotechnology today and tomorrow. AIM: How is DNA structured? Do Now: Where is DNA found and why is it important? 5/23/2017 5.1 “The DNA 200” Knowledge about DNA is increasing justice in the world. What is the most common reason why DNA analyses overturn incorrect criminal convictions? DNA is unique for each individual. Selfish dictators may owe their behaviour partly to their genes, according to a study that claims to have found a genetic link to ruthlessness. –Nature, April 2008 Whether a man has one type of gene versus another could help decide whether he’s good “husband material,” a new study suggests. –Washington Post, September 2008 The “Age of the Gene” Take-home message 5.1 DNA is a molecule that all living organisms carry in every cell in their body. Take-home message 5.1 Unique in virtually every person, DNA can serve as an individual identifier, left behind us as we go about our lives. This is a fact that is used increasingly to ensure greater justice in our society, such as through establishing the innocence of individuals wrongly convicted of crimes. 5.2 The DNA molecule contains instructions for the development and functioning of all living organisms. Two Important Features of DNA (1) DNA contains the instructions on how to create a body and control its growth and development. (2) The instructions encoded in the DNA molecule are passed down from parent to offspring. (Where have we recently seen this concept?) DNA “Double Helix” Nucleic acids and nucleotides Sugars, Phosphates, and Bases A, T, C, and G Base pairs All Teachers Go Crazy Which answer will base pair with the following sequence? AGTTCTCATGT 1. 2. 3. 4. AGTTCTCATGT ACATGAGAACT TCAAGAGTACA UCAAGAGUACA How does DNA replicate? Why is this an important process? DNA replication 1. DNA original strand “unzips” by breaking the hydrogen bonds between the complimentary base pairs. 2. New free floating nucleotides form new hydrogen bonds with the appropriate base pair with the help of DNA polymerase. 3. Two new identical of DNA molecules result, each with one strand of the original DNA as one side of the sugar-phospahate backbone. Take-home message 5.2 DNA is a nucleic acid, a macromolecule that stores information. It consists of individual units called nucleotides: a sugar, a phosphate group, and a nitrogen-containing base. Take-home message 5.2 DNA’s structure resembles a twisted ladder, with the sugar and phosphate groups serving as the backbones of the molecule and base pairs serving as the rungs. ACTIVITY Make your own DNA model. EXIT SLIP Answer the AIM How is DNA structured? 5.3 Genes are sections of DNA that contain instructions for making proteins. Why is DNA considered the universal code for all life on earth? The number of chromosomes varies from species to species. Corn has 10 unique chromosomes. Fruit flies have only four. Dogs and chickens have 39 different chromosomes. Goldfish have 47 chromosomes. Individuals in each of these species inherit one copy of each chromosome from each parent. Genes A sequence of bases in a DNA molecule that carries the information necessary for producing a functional product, usually a protein molecule or RNA Different people can have free or attached earlobes. The DNA that encodes for making free or attached earlobes is called a(n) ________, and there are two different versions of it, called __________. 1. 2. 3. 4. allele; genes trait; alleles gene; trait gene; alleles Take-home message 5.3 DNA is a universal language that provides the instructions for building all the structures of all living organisms. The full set of DNA an organism carries is called its genome. Take-home message 5.3 In prokaryotes, the DNA occurs in circular pieces. In eukaryotes, the genome is divided among smaller, linear strands of DNA called chromosomes. Take-home message 5.3 A gene is a sequence of bases in a DNA molecule that carries the information necessary for producing a functional product, usually a protein molecule or RNA. Take-home message 5.4 Only a small fraction of the DNA in eukaryotic species codes for genes. The function of the rest is still a mystery. 5.5 How do genes work? An overview Genotype • all of the genes contained in an organism Phenotype • the physical manifestations of the instructions Which molecule acts as a “middle man” between the nucleus, where transcription occurs, and the cytoplasm, where translation occurs? 1. 2. 3. 4. DNA mRNA Protein Choices 1 and 3 are correct. Take-home message 5.5 The genes in strands of DNA are a storehouse of information, an instruction book. Take-home message 5.5 The process by which this information is used to build an organism occurs in two main steps: transcription, in which a copy of the a gene’s base sequence is made, and translation, in which that copy is used to direct the production of a protein. 5.6 Transcription: reading the information coded in DNA Classroom Catalyst Transcription Activity Which answer shows the correct orientation of the mRNA sequence (Phosphate (5’) Oxygen (3’)) given the DNA sequence below? 5’-TCATCTATCT-3’ 1. 2. 3. 4. 5’-UCAUCUAUCU-3’ 5’-AGUAGAUAGA-3’ 5’-AGAUAGAUGA-3’ 5’-AGATAGATGA-3’ Take-home message 5.6 Transcription is the first step in the two-step process by which DNA directs the synthesis of proteins. In transcription, a single copy of one specific gene within the DNA is made, in the form of a molecule of mRNA, which moves where it can be translated into a protein. AIM: How are protein synthesized? Do Now: What determines the function of an enzyme? 5.7 Translation: using information from DNA to build usable molecules Several ingredients must be present in the cytoplasm for translation to occur. Free amino acids Ribosomal Transfer units RNA Classroom Catalyst Translation Activity Translate the following mRNA: 5’-CGAGCAUGGGCUUUAGG... 1. 2. 3. 4. MET-GLY-PHE-ARG… ARG-ALA-TRP-ALA-LEU… GLY-PHE-ARG-VAL-ARG… ARG-PHE-GLY-MET-SER… Take-home message 5.7 Translation is the second step in the twostep process by which DNA directs the synthesis of proteins. In translation, the information from a gene that has been carried by the nucleotide sequence of an mRNA is read, and ingredients present in the cell’s cytoplasm are used to produce a protein. 5.8 Causes and effects of mutation Alteration DNA of the sequence of bases in • can lead to changes in the structure and function of the proteins produced • can have a range of effects Breast Cancer in Humans Two human genes, called BRCA1 and BRCA2 More than 200 different changes in the DNA sequences of these genes have been detected, each of which results in an increased risk of developing breast cancer. Mutations Bad reputation Tend to be disruptive Very, very rare Which answer shows the mRNA transcribed from the DNA sequence below? 5’-TTA TCC TTT ACT CAT-3’ 1. 2. 3. 4. 5’-AUG AGU AAA GGA UAA-3’ 5’-AAU-AGG-AAA-UGA-GUA-3’ 5’- TTA-TCC-TTT-ACT-CAT-3’ 5’-UUA-AGG-AAA-TGA-GUA-3’ Which of the following changes (base substitutions and insertions are underlined and in bold) would have the most severe effect on the protein made from the mRNA in the last question? 5’-AUG AGU AAA GGA UAA-3’ 1. 2. 3. 4. AUG-AGU-AAG-GGA-UAA AUG-AGU-AAA-GGA-UAG AUG-GUA-AAG-GAU-AA AUG-AGU-AAA-GGA-GUA-A Why is it dangerous around the core of a nuclear power plant? Chemical-induced mutations Take-home message 5.8 Mutations are alterations in a single base or changes in large segments of DNA that include several genes. They are rare and when they occur in a gene usually disrupt normal physiological functioning. They play an important role in evolution. From mutation to illness in just four steps: (1) A mutated gene codes for a nonfunctioning protein, usually an enzyme. (2) The non-functioning enzyme can’t catalyze the reaction as it normally would, bringing it to a halt. From mutation to illness in just four steps: (3) The molecule with which the enzyme would have reacted accumulates, like a blocked assembly line. (4) The accumulating chemical causes sickness and/or death. Why can one change in a nucleotide sequence of a gene cause a nonfunctional protein to be produced? 1. The change can cause a stop codon to occur early in translation and the full length protein is not translated. 2. A substitution can result in one incorrect amino acid being present in the protein which can affect the shape of the protein and therefore its function. 3. An insertion or deletion can occur early in the gene and result in an incorrect translation of the protein. 4. All of the above. Take-home message 5.9 Most genetic diseases result from individual mutations that cause a gene to produce a non-functioning enzyme, which in turn blocks the functioning of a metabolic pathway. 5.10 What is biotechnology? Genetic Engineering Adding, deleting, or transplanting genes from one organism to another, to alter the organisms in useful ways Biotech advances in human health fall into three categories: (1) producing medicines to treat diseases (2) curing diseases (3) preventing diseases from occurring in the first place. If you were trying to identify the gene for insulin, which particular step of the procedure outlined below would you use? 1. Cut human DNA into small pieces with a restriction enzyme. 2. Amplify the DNA pieces with PCR. 3. Insert the gene into a plasmid and insert into bacteria. 4. Probe a gene library. Take-home message 5.10 Biotechnology is the use of technology to modify organisms, cells, and their molecules to achieve practical benefits. Take-home message 5.10 Modern molecular methods make it possible to cut and copy DNA from one organism and deliver it to another. Biotechnology has achieved some important successes in medicine, agriculture, and forensics. 5.11 Treating diseases and producing medicines with biotechnology cure diseases treating diseases • The treatment of diabetes Why do some bacteria produce human insulin? Recombinant DNA technology Several important achievements followed the development of insulin-producing bacteria, including: (1) Human growth hormone (HGH) (2) Erythropoietin Take-home message 5.11 Biotechnology has led to some notable successes in treating diseases, usually by producing medicines more efficiently and more effectively than they can be produced with traditional methods. 5.12 Curing diseases with biotechnology Gene therapy and the correction of malfunctioning genes Stem Cells Cells that have the ability to develop into any type of cell in the body Why has gene therapy had such a poor record of success in curing diseases? Gene Therapy Difficulties (1) Difficulty getting the working gene into the specific cells where it is needed. Gene Therapy Difficulties (2) Difficulty getting the working gene into enough cells and at the right rate to have a physiological effect. (3) Problems with the transfer organism getting into unintended cells. Take-home message 5.12 Gene therapy has had a poor record of success in curing human diseases. This stems primarily from technical difficulties in transferring normalfunctioning genes into the cells of a person with a genetic disease. 5.13 Preventing diseases with biotechnology Intervening to prevent diseases using biotechnology focuses on answering questions at three different points in time: I would want to know if my genome or that of a loved one carries one or more copies of a gene that can cause a genetic disease. 1. 2. 3. 4. 5. Strongly agree Agree Neutral Disagree Strongly disagree (1) Is a given set of parents likely to produce a baby with a genetic disease? (2) Will a baby be born with a genetic disease? cystic fibrosis sickle-cell anemia Down syndrome others (3) Is an individual likely to develop a genetic disease later in life? breast cancer prostate skin cancer cancer Ethical Dilemmas Discrimination Health How insurance to proceed with the information? Take-home message 5.13 Biotechnology allows us to identify: • whether a given set of parents is likely to produce a baby with a genetic disease, • whether a baby is likely to be born with a genetic disease, • and whether an individual carriers certain disease-causing genes that may have their effect later in life. Take-home message 5.13 These tools can help us to reduce suffering and the incidence of diseases, but they also come with significant potential costs, particularly the risk of discrimination. 5.14 Producing more nutritious and better food with biotechnology How might a genetically modified plant help 500 million malnourished people? Nutrient-rich “golden rice” Take-home message 5.14 Biotechnology has led to important improvements in agriculture by using transgenic plants and animals to produce more nutritious food. 5.15 More efficient and eco-friendly farming with biotechnology And he gave it for his opinion, ‘that whoever could make two ears of corn, or two blades of grass, to grow upon a spot of ground where only one grew before, would deserve better of mankind, and do more essential service to his country, than the whole race of politicians put together.’ —Jonathan Swift, Gulliver’s Travels, 1726 Almost everyone in the U.S. consumes genetically modified foods regularly without knowing it. What foods are responsible for this? Insect Resistance How can genetically modified plants lead to reduced pesticide use by farmers? Herbicide Resistance Faster Growth and Bigger Bodies Take-home message 5.15 There has been a revolution through biotechnology. Herbicide-resistant and insect-resistant crops help reduce the environmental and financial costs of producing food. 5.16 Fears and risks: the safety of genetically modified foods Fear #1. Organisms that we want to kill may become invincible. Fear #2. Organisms that we don’t want to kill may be killed inadvertently. Fear #3. Genetically modified crops are not tested or regulated adequately. Fear #4. Eating genetically modified foods is dangerous. Fear #5. Loss of genetic diversity among crop plants is risky. Fear #6. Hidden costs may reduce the financial advantages of genetically modified crops. Which food below does not potentially contain GMOs? 1. 2. 3. 4. 5. Nacho chips Fruit juice Corn flakes Rice All of the above. Take-home message 5.16 More and more genetically modified foods are being created using modern methods of recombinant DNA technology. Some legitimate fears among the public remain, however, as to the safety of these foods given that their development relies on such new technology. 5.17 DNA as an individual identifier: the uses and abuses of DNA fingerprinting What is a DNA fingerprint? Using the DNA fingerprint information below, determine which suspect was present at the crime scene? 1. 2. 3. 4. Suspect #1 Suspect #2 Suspect #3 All of the above. Crime Scene Suspect #1 Suspect Suspect #2 #3 Take-home message 5.17 Comparisons of highly variable DNA regions have forensic value in identifying tissue specimens and determining the individual from whom they came. 5.18 DNA sequences reveal evolutionary relatedness (1) Mapping Genetic Landscapes: The Human Genome Project (2) Building Earth’s Family Tree When we say humans and chimps are genetically 96% identical, what do we mean? Take-home message 5.18 Comparisons of sequence similarities across species reveal the evolutionary relatedness and make it possible to construct detailed evolutionary trees. 5.19 The promise and perils of cloning From organs to individuals Are there any medical justifications for cloning? Is it possible to clone a dinosaur? How could it be done? Take-home message 5.19 Cloning of individuals has potential benefits in agriculture and medicine, but ethical questions linger. We should pursue cloning in animals but not humans. 1. 2. 3. 4. 5. Strongly agree Agree Neutral Disagree Strongly disagree We should pursue cloning in humans for the purpose of developing or performing medical treatments. 1. 2. 3. 4. 5. Strongly agree Agree Neutral Disagree Strongly disagree