1 / 29

Exam 2 M 10/29 at 7-8:30pm in UTC 2.102A Review Th 10/25 at 5-7pm in WRW 102 Bonus #1 due now

Exam 2 M 10/29 at 7-8:30pm in UTC 2.102A Review Th 10/25 at 5-7pm in WRW 102 Bonus #1 due now Last day to drop with a ‘Q’…10/24. Genetic Engineering: Direct manipulation of DNA. Bacteria can be modified or serve as intermediates. a typical bacteria. Bacterial DNA. plasmid DNA.

tsimon
Download Presentation

Exam 2 M 10/29 at 7-8:30pm in UTC 2.102A Review Th 10/25 at 5-7pm in WRW 102 Bonus #1 due now

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Exam 2 M 10/29 at 7-8:30pm in UTC 2.102A Review Th 10/25 at 5-7pm in WRW 102 Bonus #1 due now Last day to drop with a ‘Q’…10/24

  2. Genetic Engineering: Direct manipulation of DNA

  3. Bacteria can be modified or serve as intermediates

  4. a typical bacteria Bacterial DNA plasmid DNA

  5. A typical bacterial plasmid used for genetic engineering

  6. Moving a gene into bacteria via a plasmid

  7. What problems exist for expressing eukaryotic gene in bacteria? Bacterial DNA plasmid DNA

  8. Reverse transcriptase can be used to obtain coding regions without introns.

  9. After RT, PCR will amplify the gene or DNA Fig 20.14

  10. Moving a gene into bacteria via a plasmid RT and PCR

  11. Restriction Enzymes cut DNA at specific sequences

  12. Restriction enzymes cut DNA at a specific sequence

  13. Cutting the plasmid and insert with the same restriction enzyme makes matching sticky ends Fig 20.2

  14. A typical bacterial plasmid used for genetic engineering

  15. Using sticky ends to add DNA to a bacterial plasmid Fig 20.2

  16. If the same restriction enzyme is used for both sides, the plasmid is likely to religate to itself. Fig 20.2

  17. The plasmid is treated with phosphatase to remove the 5’-P, preventing self-ligation Fig 20.2

  18. Transformation of bacteria can happen via several different methods. Fig 20.8

  19. Bacteria can take up DNA from the environment Fig 7.2

  20. Transformation of bacteria can happen via several different methods all involving perturbing the bacterial membrane: • Electroporation • Heat shock • Osmotic Stress Fig 20.8

  21. How can you know which bacteria have been transformed, and whether they have the insert?

  22. Figure 20-5 Resistance genes allow bacteria with the plasmid to be selected. Fig 20.5 Bacteria with the resistance gene will survive when grown in the presence of antibiotic

  23. Figure 20-5 Is the insert present?Plasmids with the MCS in the lacZ gene can be used for blue/white screening… Fig 20.5

  24. A typical bacterial plasmid used for genetic engineering

  25. Figure 20-5 Intact lacZ makes a blue color when expressed and provided X-galactose Fig 20.5

  26. Figure 20-5 When the lacZ gene is disrupted, the bacteria appear white Fig 20.5

  27. Blue/white screening: Transformed bacteria plated on antibiotic and X-gal plates.Each colony represents millions of clones of one transformed cell.

  28. Successful transformation will grow a colony of genetically modified bacteria Fig 20.4

  29. RT and/or PCR Inserting a gene into a bacterial plasmid

More Related