F215 OCR Genetic Engineering

?
View mindmap
  • Genetic Engineering
    • eg Insulin & Golden Rice
    • 1. The Required Gene Is Obtained
      • - Use mRNA (Avoids finding gene amongst entire human genome
        • - From B cells in the IOL. Transcribe and Translate the Human Insulin Gene
          • - Isolate mRNA & add reverse Transcriptase= produces cDNA which is single stranded
            • - Add DNA Polymerase & free nucleotides to join SP Backbone add DNA Ligase (phosphodiester bond) to form DNA
      • Use Restriction enzymes & find with gene probe
    • 2. A copy of the gene is placed into a Vector
      • What is a Vector? A vehicle that carries the gene of interest into target cell
      • Placed in Plasmid, small circular loop of DNA with antibiotic resistance that can self replicate
        • Cut open using Restriction Enzymes, create sticky end that are complementary to insulin gene
          • Add artificial sticky ends onto insulin gene
            • H Bonds form between complementary bases= 2 A&T, 3 G&C
              • RecombinantPlasmid formed = Joining of DNA from 2 different species
    • 3. The vector carries gene into Recipient Cell = E.Coli
      • Bacteria grow on agar plate don't know which ones have taken it up so we use Marker Genes
        • Genes that allow you to identify & select bacteria that have taken up the gene of interest eg insulin
          • Plasmids have two different genes Ampicillin & Tetracycline
            • Insulin in the vector breaks the tetracycline gene so no longer resistant but still resistant to ampicillin
        • Process = Replica Plating. Ampicillin agar all bacteria grow then transfered to Tertracycline agar only plasmid without insulin will grow
          • By keeping track of the colonies the desirable colonies can be identified
    • 4. Transformed bacteria contaning insulin gene are grown on a large scale in a fermenter
      • Bacteria is transcribed and translated and insulin polypeptide is produced then extracted and purified
      • Protein Synthesis
        • Bacteria is transcribed and translated and insulin polypeptide is produced then extracted and purified

Comments

ellielewis98

Report

Click Edit Resource to be able to see it properly as its too big to fit in the screen so it overlaps

Similar Biology resources:

See all Biology resources »See all DNA, genetics and evolution resources »