Chapter 13 Genetic Engineering

1. Chapter 13Genetic Engineering Section 13-2Manipulating DNA

2. Manipulating DNA Key Concept: Scientists Use Their Knowledge Of The Structure of DNA And Its Chemical Properties To Study and Make Changes To DNA Molecules

3. Manipulating DNA Key Concept (cont.) Different Techniques are used to Extract DNA from Cells, to Cut DNA into Smaller Pieces, to Identify the Sequence of Bases in the DNA Molecule, and to Make Unlimited Copies of DNA

4. The Tools of Molecular Biology Genetic Engineering The Process of Making Changes In The DNA Code of a Living Organism

5. The Tools of Molecular Biology DNA Extraction Use Chemicals To Lyse Cells Lysis: a process of disintegration or dissolution (as of cells)

6. The Tools of Molecular Biology Cutting DNA DNA Too Large To Work With, So It Is Cut Up Using: Restriction Enzymes: • They Are Very Precise • 100’s Are Known • Each One Cuts DNA At Specific Sequence of Nucleotides

7. Restriction Enzymes

8. The Tools of Molecular Biology Separating DNA Gel Electrophoresis • Mixture of Fragments Put In One End of Gel • Electric Voltage Applied • Fragments Travel Toward Positive End of Gel

9. The Tools of Molecular Biology Gel Electrophoresis (cont.) • Smaller The Fragment, The Faster It Moves • Used To: • Compare Genomes • Locate Individual Genes • Identify Base Pair Sequence

10. Gel Electrophoresis

11. Gel Electrophoresis

12. Gel Electrophoresis

13. Gel Electrophoresis

14. Using The DNA Sequence Once The DNA Is A Manageable Size, You Can: • Read The Nucleotide Sequences • Modify The Genome

15. Using The DNA Sequence Reading The Sequences • Now Automated • Small, Single Strands of DNA • Add Enzyme That Makes Complementary Strand • Add Nucleotides, Some That Are Labeled With A Specific Color of Fluorescent Dye

16. Using The DNA Sequence

17. Using The DNA Sequence

18. Using The DNA Sequence Reading The Sequences (cont.) • Addition of a Labeled Base To Strand Terminates Replication • Produces Multiple, Labeled Strands of Different Lengths • Each Terminal Base Is Color Coded

19. Using The DNA Sequence Reading The Sequences (cont.) • Separate by Electrophoresis • Read The Base Sequences In Order, By Color Codes

20. Using The DNA Sequence

22. Automated Sequencing

23. Changing The DNA Sequence Cutting & Pasting You Can Take A Gene From One Organism & Attach It To The Gene of Another Organism

24. Changing The DNA Sequence Cutting & Pasting (cont.) Mixing Genes From Different Organisms Results In Recombinant DNA

25. Using The DNA Sequence Making Copies PCR = Polymerase Chain Reaction Quickly Makes Multiple Copies Of Small DNA Targets

26. PCRPolymerase Chain Reaction Add Short Strand of Complementary DNA To Each End Of The DNA You Want To Copy Primers Provide Location For DNA Polymerase To Attach

27. PCRPolymerase Chain Reaction • Heat To Separate The Strands Of DNA • Cool To Allow Primers To Attach The Primers • Increase Temperature To Activate TAQ Polymerase (High Temperature DNA Polymerase) • Repeat 20 – 30 Times • Copies Also Act As Templates

28. PCRPolymerase Chain Reaction Within A Few Hours You Will Have Millions of Copies

29. PCRPolymerase Chain Reaction

30. PCRPolymerase Chain Reaction • Kary Mullis • Invented PCR • Source of High TemperatureDNA Polymerase (taq Polymerase) • Bacteria In hot springs of Yellowstone Park

31. PCRPolymerase Chain Reaction Yellowstone HotspringSource of High Temperature PCR DNA Polymerase(taq Polymerase)