2. Biotechnology Biology- Regular John Murnan Etowah High School

3. Welcome to Biotech Inc.

4. Your new job requires some tools Biotech Inc. does research into developing products for Genetic Engineering, DNA fingerprinting and Cloning As interns you will learn how to use some new tools and techniques Please open your tool box and lets see what’s inside it.

5. Restriction Enzymes –Act like scissors –Cut DNA at specific places

6. Recognition sites of different restriction enzymes

7. Restriction site function

8. Function of restriction enzymes

9. Ligase Acts like glue Connects two pieces of DNA –Makes bond between sugar-phosphate on backbone.

10. DNA Polymerase Acts like a copier –Copies a strand of DNA Requires free nucleotides to make the copies

11. Retroviruses Can insert DNA into a chromosome in a living cell –HIV, the virus that causes AIDS is one type.

12. Some smaller tools Very thin syringe –Used to remove nucleus from cells Micropipette –A very small eyedropper –Used to move very small amounts of fluid

13. Gel Electrophoresis Sorts molecules by size –Uses a gel like jello and an electric current.

14. Gel Electrophoresis Setup

15. Gel Electrophoresis

16. Pictures of gel electrophoresis

17. PCR Uses DNA Polymerase and free nucleotides to make millions of copies of DNA from a single strand.

18. Overview of PCR

19. Effect of PCR

20. Genetic Engineering

21. Manipulating organisms to create products Use restriction enzymes, ligase, Gel Electrophoresis and PCR Inserts foreign genes into bacteria –Makes bacteria produce proteins –Bacteria can be used to store genes

22. Plasmid Ligation Puts DNA from humans (or other species) into bacteria DNA.

23. Review of Bacteria Structure Nucleoid Region- is bacteria chromosome PLASMID- small rings of DNA that have only a few genes. Plasmid

24. Ligation of plasmid 1

25. Ligation of plasmid 2

26. Genetic Engineering with plasmid

27. Possible products of genetic engineering Using Plasmid Ligation techniques, we can make both proteins and genes that can be used to

28. DNA Fingerprinting

29. Forensic Scientists Use restriction enzymes, Gel Electrophoresis, PCR Can identify any person based on their DNA sequences. Each of us has unique sequences.

30. What is an RFLP? Sometimes mutations occur which cause recognition sites to be changed. If a person has a mutation, then the site is changed and DNA of different lengths is produced using the same restriction enzyme. Restriction Fragment Length Polymorphism

31. Why you are different than other people

32. Southern Blot 1

34. Southern blot results

35. DNA fingerprinting example

36. http://www.koshlandsciencemuseum.org/ex hibitdna/index.jsphttp://www.koshlandsciencemuseum.org/ex hibitdna/index.jsp

37. Cloning

38. Cloning Activity http://gslc.genetics.utah.edu/units/cloning/cl ickandclone/http://gslc.genetics.utah.edu/units/cloning/cl ickandclone/

39. Ethics of cloning Occurs naturallly (identical twins) Easier screening of embryos for genetic diseases Increase chances for infertile couples to have children High risk of developmental errors leading to miscarriages Cells are as old as the individual they are taken from

40. Gene Therapy

41. Gene therapy example

42. Gene therapy vectors

43. Delivery methods

44. Gene Therapy example