CAS BI 105 Lecture Notes - Lecture 18: Polymerase Chain Reaction, Restriction Enzyme, Restriction Site
30 Apr 2018
School
Department
Course
Professor
Biotechnology
Biotechnology
•Modifying organisms/cells/molecules of cells in order to achieve some result
•Genetic Engineering - manipulation of the genetic material of an organism
•Recombinant DNA Technology - “recombining” genes of different organisms to create an organism
with new capabilities
•Using genetic information for diagnostics/treatment/crime-solving
Impacts (do not need to memorize)
•Human health impacts
•Producing new medicine
•Treating inherited diseases
•Preventing genetically transferred diseases
•Agriculture impacts
•Improving the taste/nutrition of food
•Increasing the efficiency of farming
•Decrease the ecological impact of farming
•Crime & Diagnostics impacts
•Leads to new information about a person’s actions/location
•Leads to information about a person’s susceptibility to disease
Recombinant DNA Technology
•*Most of our drugs are produced in bacteria
•Steps -
•1. Chop DNA from its donor species
•2. Amplify the single copy of DNA into many copies
•3. Insert the DNA into bacteria
•4. Grow the bacteria and send it through transcription/translation to produce proteins
•5. Harvest the ^protein and purify it to be used in humans
Cutting DNA
•Bacteria use enzymes (restriction enzymes) to protect themselves from viruses
•Restriction Enzymes recognizes specific sequences of DNA and splice at these points
•^scientists use these enzymes to cut DNA
•Restriction Enzymes - cuts double-stranded DNA (dsDNA) at specific sequences (“restriction site”)
•^cuts at the same place on both strands
Amplify DNA
•In order to make many copies of a DNA stretch, it would involved many cells and a lot of time
•^Polymerase Chain Reaction accomplishes this much faster and efficiently
Recombination in Eukaryotes
•Bacteria take up DNA from the environment and replicate it on its own OR integrate it into its 1
chromosome
•Barriers to getting external DNA to be expressed in a eukaryotic cell?
•Nuclear Membrane, Cell wall
•Ways to overcome gene delivery problems in eukaryotes?
•Viruses
Recombination in DNA
•How do we get recombined DNA into an organism? How do we know if the organism is using newly
inserted genes?
•Getting in -
find more resources at oneclass.com
find more resources at oneclass.com
Document Summary
Recombinant dna technology: *most of our drugs are produced in bacteria, steps , 1. Chop dna from its donor species: 2. Amplify the single copy of dna into many copies: 3. Grow the bacteria and send it through transcription/translation to produce proteins: 5. Harvest the ^protein and purify it to be used in humans. Amplify dna: in order to make many copies of a dna stretch, it would involved many cells and a lot of time, ^polymerase chain reaction accomplishes this much faster and efficiently. Recombinant dna - are you there: many variations, ex: antibiotic resistance, include ampicillin (type of antibiotic) resistance in plasmid, plate bacteria in the presence of ampicillin. Recombinant dna technology: the first type of technology created insulin from bacteria. Analyzing genomes: purpose = to look for diseases, complementary binding - to search a genome for a mutated disease/disease risk factor, single gene diseases (ex: pku, cystic fibrosis, single gene risk factors (ex: brca1/bcra2)
