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IB Genetics Topic 4:. Genetic Engineering. Topics in Genetic Engineering. Polymerase Chain Reaction (PCR) Gel electrophoresis DNA profiling The genome project Gene transfer Genetic modification in plants and animals Cloning. We will be brief!.
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IB Genetics Topic 4: Genetic Engineering
Topics in Genetic Engineering • Polymerase Chain Reaction (PCR) • Gel electrophoresis • DNA profiling • The genome project • Gene transfer • Genetic modification in plants and animals • Cloning
We will be brief! • The Blog has videos and games for you to cement your understanding of each topic!
4.4.1: The Polymerase Chain Reaction • Kary Mullis, 1984, Nobel Prize 1993 • ‘Reviewers and Biographers often latch on to his enjoyment of drugs, womanising and surfing to paint him as some sort of cool rock star rebel…but…’
What does PCR do? • It quickly amplifies a tiny sample of DNA into millions of identical copies • This produces enough DNA to allow further analysis and study.
The DNA sequences amplified by PCR are used for: • Forensic identification - crime (CSI….) • Paternity suits • The Genome project: determination of the entire human genetic code • Diagnosing diseases and genetic disorders – genotype analysis • Detection of bacteria and viruses in the environment • Evolutionary research
How does PCR work? First, you need to extract DNA from cells. Next, PCR requires: • Custom-made DNA primers • (TAQ) DNA polymerase • Nucleotides for complementary base pairing • HEATING and COOLING: Thermocycler machine PCR can produce 100 billion copies of the DNA sample in a few hours
Let’s do it! • PCR the movie • PCR the cartoon • PCR the game
4.4.2: Gel electrophoresis • Commonly the DNA sample is first amplified by PCR • The DNA sample is chopped into fragments using restriction enzymes • Gel electrophoresis separates fragments of DNAaccording to their size • DNA samples are placed in a gel, fluorescent marker added (tag attached to a triplet), and an electric charge is applied to push fragments along • The shortest fragments travel furthest through the gel, while the longest (heaviest) remain closer to their origin • This lets us ‘match’ the sample against another DNA sample or standard
Let’s do it! • Virtual Lab 1 • Virtual lab
4.4.3: DNA profiling HOW? APPLICATIONS Numerous techniques for DNA profiling: Criminal forensics Paternity suits Ecology – identifying relationships in migrating birds, whales etc Evolutionary genetics • DNA sample (blood, semen, cheek swab) • Sample is amplified by PCR, then matched against known samples using gel electrophoresis • We look for standard tandem repeating DNA: highly repetitive sequences from non-coding region of DNA • STR’s are unique to individuals
Famous examples of DNA profiling • The innocence project • The famous blue dress • Identifying the Mona Lisa • Identifying the Romanovs
DNA profiling: paternity and forensics • The fluorescent banding pattern shows up for each DNA fragment and can be compared with test/ known sample • Usually >4 ‘STR’ sites are used to establish paternity or identity
4.4.6: The Human Genome project We’re all the same! We’re all different! Welcome to the human genome
Started in 1990, first draft published in 2003, ongoing refinement… Amazing international effort • The BBC tell us about the genome 10 years on • Hank tells us some surprises about the human genome project • ..so what has the human genome project achieved, so far?
Key outcomes of the Human Genome project • We identified the number (30,000) and loci of all the genes of our genome 1. Human health: Medical diagnostics, treatments, pharmacogenomics, gene therapy • We identified many new proteins and their functions 2. Transgenics: move production of beneficial proteins from one species to another, design novel proteins • We compared human DNA with that of other species 3. Evolutionary genetics and evolutionary history 4. Bio-informatics: Genetic databases and the bio-informatics industry
The HGP and human health • 2 out of 3 of us will die of a genetically related disease • Many (some say all) diseases have a genetic cause • Genetic report cards can identify ‘potential risk’ of future illnesses • The future is here...
Do you want to know your genome? Do you want to know your genome? Are you sure?
The human genome and new medicine (1): Gene therapy • Find defective genes and ‘fix them’ • Insert a healthy normal gene or gene product (lipid, protein)to achieve normal function • Most effective for single gene disorders: cystic fibrosis, Sickle Cell disease, • gene therapy in depth • A lucrative business...
The human genome provides avenues for new medicines • Identify beneficial molecules which are produced in healthy people • Identify the gene which produces the desirable molecule • Copy that gene and use it as a recipe to synthesise the molecule in a laboratory (or in another organism!) • Distribute the beneficial molecule as a new treatment
The HGP and medicine (3): Pharmacogenomics Uses information about your genetic make-up to determine the drug, and drug doses, that will work best for you Currently used for treatment of: • HIV • Breast cancer • Colon cancer • Mental illness more info here...
3. Using the HGP to understand human evolution By sequencing and databasing genes, we can see similarities and differences between species • The closer the genome match, the closer their evolutionary history • Human Chromosome 2 came from fusion of two great ape chromosomes • Karl Miller on human evolution • The time-tree of evolution
Ethics and the human genome How companies are patenting your genetic code
4.4.8: GENETIC ENGINEERING, (MODIFICATION, TRANSGENICS)BIOTECHNOLOGY
The basic premise for genetic engineering… The genetic code is universal • All living things share the same code • Each codon codes for the same animoaacid, regardless of the species • Genes can be transferred from species to species, to produce the same product We can cut, copy and paste DNA between species
Gene transfer 101: the basics of cutting, copying, pasting and cloning genes • An introduction • Step-by-step
How does genetic engineering work? • Cut out desired DNA using restriction endonuclease • Prepare plasmid using the same restriction enzyme • Insert DNA into plasmid using DNA ligase • Insert plasmid (vector) into host cell
Applied gene transfer: Gene Therapy Gene therapy for treatment of Severe Combined Immunodeficiency Disease (2)
Genetic Engineering: Making Human insulin • E. Coli make human insulin
Genetic Engineering: Making Factor IX in sheep… 1997: Factor IX isolated and purified from sheep’s milk to treat one hereditary form of haemophilia (Haemophilia B, Christmas Disease)
Transgenic plants: The controversies… The controversies….
Glowing animals?....what’s the big deal? Worth a Nobel prize….
Further Applications of Transgenic Technology • Vaccines (transgenic yeast produces vaccine for Hepatitis B) • Transgenic mice widely used for human disease research • Transgenic male mosquitoes carrying ‘lethal gene’ used to reduce the incidence of Dengue fever
Genetic modification: the PROs… • GM crops should improve yields, quality and food security • Pest-resistant crops will reduce the need for pesticides • GM can produce crops which provide dietary supplements such as retinol • GMO’s used to produce rare proteins for medicine or vaccines will be cheaper and less polluting than conventional methods • GM allows farmers greater control for selective breeding of crops and animals
Genetic modification: the CONs… • No one knows the long-term effects of GMO’s in the biosphere PRECAUTIONARY PRINCIPLE • Genes from GM crops are easily integrated into the wild type crop • Genes may be able to cross species • Crops which produce toxins to kill insects could prove toxic to humans • Large portions of the human food supply may fall under the control of a small number of companies • High-tech agricultural ones are not necessarily better than simpler solutions • A proliferation of GMO’s could drastically reduce natural biodiversity
What is a clone? • A group of genetically identical organisms • A group of cells artificially derived from a single parent