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Manipulating DNA PowerPoint Presentation

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Description : Check out this medical PowerPoint presentation titled "Manipulating DNA".
This medical PowerPoint presentation talks about Manipulating DNA, the process of modifying the genetic material of an organism.
There are several techniques used for DNA manipulation, including Restriction enzymes, Polymerase chain reaction (PCR), DNA sequencing, Gene editing, and DNA cloning.
DNA manipulation is an important tool in genetics and biotechnology. It can be used to study gene function, develop new therapies for genetic diseases, and create genetically modified organisms with desirable traits. However, there are also ethical concerns associated with DNA manipulation, particularly when it comes to creating genetically modified organisms.

Tags : Manipulating DNA, Restriction enzymes, Polymerase chain reaction (PCR), DNA sequencing, Gene editing, DNA cloning, Genetics, Genetics PowerPoint presentation

Published on : Mar 27, 2023 | Views : 1544 | Downloads : 1

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KEY CONCEPT Biotechnology relies on cutting DNA at specific places.
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What steps does DNA Technology Involve?
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Scientists use several techniques to manipulate DNA. Chemicals, computers, and bacteria are used to work with DNA. Scientists use these tools in genetics research and biotechnology.
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Restriction enzymes cut DNA. Restriction enzymes act as “molecular scissors.” come from various types of bacteria allow scientists to more easily study and manipulate genes cut DNA at a specific nucleotide sequence called a restriction site
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Different restriction enzymes cut DNA in different ways. each enzyme has a different restriction site
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some cut straight across and leave “blunt ends” some make staggered cuts and leave “sticky ends”
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Restriction maps show the lengths of DNA fragments. Gel electrophoresis is used to separate DNA fragments by size. A DNA sample is cut with restriction enzymes. Electrical current pulls DNA fragments through a gel.
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Smaller fragments move faster and travel farther than larger fragments. Fragments of different sizes appear as bands on the gel.
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A restriction map shows the lengths of DNA fragments between restriction sites. only indicate size, not DNA sequence useful in genetic engineering used to study mutations DNA Electrophoresis Animation
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Examples When is the technique used to separate DNA, To get a DNA fingerprint for forensic purposes To get a DNA fingerprint for paternity testing To get a DNA fingerprint so that you can look for evolutionary relationships among organisms When is the technique used to separate proteins Many know you can separate DNA fragments – thank you TV and movies – but you can also separate proteins if the correct gel is used Think about it – do all proteins contain the same number of amino acids?
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Disease example Testing for DNA fragments that indicate having IBS Only find fragments in people who have the disease – lanes 2-5 Lanes 6-9 do not have the disease What does it mean that land 2 has IBS but no band?
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Myofibrils prepared from white skeletal muscle of rabbit contain all the myofibrillar proteins arranged in complex structures described above. The individual myofibrillar proteins and, moreover, the individual subunits of the proteins will be separated in the gel based on their molecular mass. Separation of proteins in a muscle fiber
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KEY CONCEPT The polymerase chain reaction rapidly copies segments of DNA.
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PCR uses polymerases to copy DNA segments. PCR makes many copies of a specific DNA sequence in a few hours. PCR amplifies DNA samples. PCR is similar to DNA replication.
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PCR is a three-step process. PCR uses four materials. DNA to be copied DNA polymerase A, T, C, and G nucleotides two primers
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The three steps of PCR occur in a cycle. heat is used to separate double-stranded DNA molecules primers bind to each DNA strand on opposite ends of the segment to be copied DNA polymerase binds nucleotides together to form new strands of DNA
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Each PCR cycle doubles the number of DNA molecules.
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Uses of PCR PCR can isolate a single gene our of hundreds of thousands. (just to give you an idea- a gene of 3000 bp (or less) can readily be fished out of a human genome of 3 billion bp) PCR can also be used to introduce mutations into genes, which can then be studied to understand the effects of these mutations on the function of the encoded protein. Since it is possible to amplify large amounts of DNA from tiny traces, it is used by forensic labs to get sufficient DNA from evidence at crime scenes to help identify "whodunnit". For the same reason as above, PCR can be used to detect the presence of infectious agents,for example, the AIDS virus (HIV) in blood, long before it can be detected by other meansavailable today.
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Examples Disease Tissue typing for organ transplants Prenatal testing – are you are carrier for a genetic disease Epidemics - ie strain in wildlife/human Forensics Fingerprint testing – samples to crime scene or close relative identification Research Can get known segments of genetic disease mutation Phylogenic analysis to link to ancient sources – mammoths, and neanderthals OH MY! Patterns of gene expression – ie through different developmental stages
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KEY CONCEPT DNA fingerprints identify people at the molecular level.
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A DNA fingerprint is a type of restriction map. DNA fingerprints are based on parts of an individual’s DNA that can by used for identification. based on noncoding regions of DNA noncoding regions have repeating DNA sequences number of repeats differs between people banding pattern on a gel is a DNA fingerprint
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DNA fingerprinting is used for identification. DNA fingerprinting depends on the probability of a match. Many people have thesame number ofrepeats in a certainregion of DNA. The probability that two people share identicalnumbers of repeats inseveral locations isvery small.
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Individual probabilities are multiplied to find the overall probability of two DNA fingerprints randomly matching. Several regions of DNA are used to make DNA fingerprints.
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Family with two daughters and two sons Mom had a daughter from a previous marriage What do you think the situation is with Son 2?
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DNA from crime scene (sperm DNA), victim, and, victim’s boyfriend and two suspects Ladders are used to judge the size of the DNA fragments
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DNA fingerprinting is used in several ways. evidence in criminal cases paternity tests immigration requests studying biodiversity tracking genetically modified crops Cleared by DNA list Innocence project
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Variable Number Tandem Repeats location in a genome where a short nucleotide sequence is organized as a tandem repeat. These can be found on many chromosomes, and often show variations in length between individuals
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How can it be applied? Baby 81. DNA saved the day when it helped bring parents and a lost child together after the tsunami hit Sri Lanka on the day after Christmas 2004. Lots of people were separated from their families. In particular, a lot of children were separated from their parents. One child was called "Baby 81" when he was admitted to the hospital. As you can imagine, many parents who lost children in the tsunami hoped that Baby 81 might be theirs. The hospital didn't want to give the child away to the wrong family, so a judge ordered DNA testing to prove who the parents really were. Everything turned out in the end for one family when the DNA fingerprinting tests came back and they were able to reclaim their son.
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Application #2 9/11 and the World Trade Center When the airplanes hit the World Trade Center in New York on September 11, 2001, nearly 3000 people were killed. The impact of the planes and the fires that started made it impossible to identify many of the victims. In order to give closure to the families of the people who died, the medical examiner's office in New York has been working on identifying every remain found using DNA. Even reclaiming a little bit of their loved one has been important to some families. People turned in toothbrushes and hair brushes and anything else that might have traces of their loved one's DNA on it. The medical examiner's office then tried to match the DNA in the remains to these samples. Unfortunately, they had to stop the DNA identifications in February 2005researchers had reached the limits of DNA fingerprinting technology. There are still samples that contain tiny bits of DNA, but they will have to wait until methods like the lab-on-a-chip that needs less sample become available.
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KEY CONCEPT DNA sequences of organisms can be changed.
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Entire organisms can be cloned. A clone is a genetically identical copy of a gene or of an organism.
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Cloning occurs in nature. bacteria (binary fission) some plants (from roots) some simple animals (budding, regeneration)
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Mammals can be cloned through a process called nuclear transfer. nucleus is removed from an egg cell nucleus of a cell from the animal to be cloned is implanted in the egg
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Cloning has potential benefits. organs for transplant into humans save endangered species Cloning raises concerns. low success rate ( Dolly was the only one out of 277 embyros) clones “imperfect” and less healthy than original animal decreased biodiversity
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Animals Clones Carp Sheep Mouse Cows Goats Pigs Mouflon Rhesus Monkey Guar Rabbit Cat Rat Mule Horse African Wildcat Deer Ferret Buffalo Dog Wolves
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New genes can be added to an organism’s DNA. Genetic engineering involves changing an organism’s DNA to give it new traits. Genetic engineering is based on the use of recombinant DNA. Recombinant DNA contains genes from more than one organism.
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Bacterial plasmids are often used to make recombinant DNA. plasmids are loops of DNA in bacteria restriction enzymes cut plasmid and foreign DNA foreign gene inserted into plasmid
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Genetic engineering produces organisms with new traits. A transgenic organism has one or more genes from another organism inserted into its genome.
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Transgenic bacteria can be used to produce human proteins. gene inserted into plasmid plasmid inserted into bacteria bacteria express the gene Can also create a transgenic animal in cases where bacteria aren’t possible Transgenic plants are common in agriculture. transgenic bacteriainfect a plant plant expressesforeign gene many crops are nowgenetically modified(GMO)
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Transgenic Animals???? In cases like insulin production, the gene can be inserted into a plasmid and replicated in bacteria and grown in vats Sometimes, mammalian cells are needed so they use diary animals like cows and goats – why MILK!!!! Can just milk the animal and harness large amounts of the protein produced. Gene is given an on/off switch that only turns on in mammary cells => only in milk Treatments for hemophilia, blood clotting disorders, Cystic fibrosis, and anemia are some being studied Video on how it is done
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GMOs Oh My Pros WHO, American Academy of Sciences, and AMA say they are safer than traditional methods More food Less stress on environment Better products Cons Rise of superweeds Health Problems Frankenfoods Inadvertent creation of other products due to introduction of genes and bacteria Other organisms could be harmed in an ecosystem
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Transgenic animals are used to study diseases and gene functions. transgenic mice used to study development and disease gene knockout mice used to study gene function
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Genetically modified cattle treating Ebola??? Using cloned, genetically engineered cattle – identical, part human Incubating antibodies against ebola – will be in blood plasma Genetically engineered to produce human antibodies INSTEAD of cattle antibodies Vaccinated against deadly disease to make them produce antibodies which can then be removed and used to treat humans Similar to transfusing patients with survivor serum 30-60L of plasma enough = 500-1,000 doses/month/animal Currently being used on mice – can protect if only exposed one day with one dose, beyond that working on everyday doses Next monkeys and then onto humans Hantavirus, flu, MERS as well – 50 cattle currently
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Scientists have concerns about some uses of genetic engineering. possible long-term health effects of eating GM foods possible effects of GM plants on ecosystems and biodiversity
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KEY CONCEPT Entire genomes are sequenced, studied, and compared.
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Genomics involves the study of genes, gene functions, and entire genomes. Genomics is the study of genomes. can include the sequencing of the genome comparisons of genomes within and across species
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Gene sequencing is determining the order of DNA nucleotides in genes or in genomes. The genomes of several different organisms have been sequenced.
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The Human Genome Project has sequenced all of the DNA base pairs of human chromosomes. analyzed DNA from a few people still working to identify and map human genes
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Technology allows the study and comparison of both genes and proteins. Bioinformatics is the use of computer databases to organize and analyze biological data. DNA microarrays are used to study the expression of many genes at once. Proteomics is the study and comparison of proteins.
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DNA Microarray A DNA micorarray allows scientists to perform an experiment on thousands of genes at the same time. Each spot on a microarray contains multiple identical strands of DNA. The DNA sequence on each spot is unique. Each spot represents one gene. Thousands of spots are arrayed in orderly rows and columns on a solid surface (usually glass). The precise location and sequence of each spot is recorded in a computer database. Microarrays can be the size of a microscope slide, or even smaller. http://learn.genetics.utah.edu/content/labs/microarray/
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DNA Microarray http://learn.genetics.utah.edu/content/labs/microarray/
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DNA Microarray – how it works Blood sample is taken from the patient and a normal control is uses DNA is denatured so the two strands come apart DNA is cut into fragments and each fragment is labeled with a fluorescent dye – patient green, normal red Both sets of DNA are placed on a chip and allowed to bind to the synthesized DNA on the chip IF NO MUTATION – patient and control DNA will bind to the same sequences on the chip IF PATIENT HAS A MUTATION – those sequences containing the mutation will bind to the mutated sequences on the chip
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DNA Microarray - Applications Gene Discovery: DNA Microarray technology helps in the identification of new genes, know about their functioning and expression levels under different conditions. Disease Diagnosis: DNA Microarray technology helps researchers learn more about different diseases such as heart diseases, mental illness, infectious disease and especially the study of cancer. Until recently, different types of cancer have been classified on the basis of the organs in which the tumors develop. Now, with the evolution of microarray technology, it will be possible for the researchers to further classify the types of cancer on the basis of the patterns of gene activity in the tumor cells. This will tremendously help the pharmaceutical community to develop more effective drugs as the treatment strategies will be targeted directly to the specific type of cancer.
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DNA Microarray - Applications Drug Discovery: Microarray technology has extensive application in Pharmacogenomics. Pharmacogenomics is the study of correlations between therapeutic responses to drugs and the genetic profiles of the patients. Comparative analysis of the genes from a diseased and a normal cell will help the identification of the biochemical constitution of the proteins synthesized by the diseased genes. The researchers can use this information to synthesize drugs which combat with these proteins and reduce their effect. Toxicological Research: Microarray technology provides a robust platform for the research of the impact of toxins on the cells and their passing on to the progeny.
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EXAMPLE
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KEY CONCEPT Genetics provides a basis for new medical treatments.
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Genetic screening can detect genetic disorders. Genetic screening involves the testing of DNA. determines risk of havingor passing on a geneticdisorder used to detect specificgenes or proteins can detect some genesrelated to an increasedrisk of cancer can detect some genesknown to cause geneticdisorders
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Gene therapy is the replacement of faulty genes. Gene therapy replaces defective or missing genes, or adds new genes, to treat a disease.
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Several experimental techniques are used for gene therapy. genetically engineered viruses used to “infect” a patient’s cells insert gene to stimulate immune system to attack cancer cells insert “suicide” genes into cancer cells that activate a drug
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Examples Gene therapy is being used in many ways. For example, to: Replace missing or defective genes; Deliver genes that speed the destruction of cancer cells; Supply genes that cause cancer cells to revert back to normal cells; Deliver bacterial or viral genes as a form of vaccination; Provide genes that promote or impede the growth of new tissue; and; Deliver genes that stimulate the healing of damaged tissue.
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Parkinsons Patients with Parkinson's disease gradually lose cells in the brain that produce the signaling molecule dopamine. As the disease advances, patients lose the ability to control their movements. A small group of patients with advanced Parkinson's disease were treated with a retroviral vector to introduce three genes into cells in a small area of the brain. These genes gave cells that don't normally make dopamine the ability to do so. After treatment, all of the patients in the trial had improved muscle control.
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Hemophilia People with hemophilia are missing proteins that help their blood form clots. Those with the most-severe forms of the disease can lose large amounts of blood through internal bleeding or even a minor cut. In a small trial, researchers successfully used an adeno-associated viral vector to deliver a gene for Factor IX, the missing clotting protein, to liver cells. After treatment, most of the patients made at least some Factor IX, and they had fewer bleeding incidents.
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Gene therapy has many technical challenges. inserting gene into correct cells controlling gene expression determining effect on other genes