1. Biotechnology The use of biological processes, organisms, or systems to manufacture products intended to improve the quality of human life.

2. Transgenics Genetic engineering makes it possible to transfer DNA sequences from one organism to another REMEMBER- DNA is universal A gene taken from one species can be replicated in another species.

3. Recombinant DNA Recombinant DNA contains DNA from 2 or more different sources. So how does DNA get from one species to another? Vector: something used to transfer DNA from one organism to another –Microinjection –Gene Gun –Virus –Bacteria Plasmid

4. Bacteria as vectors plasmidsBacteria are used because they have plasmids A plasmid is a circular piece of DNA that exists apart from the chromosome and replicates independently of it.

5. So how do I take a gene from one organism and put it into another? We have to cut it using a R estriction E nzyme

6. What are Restriction Enzymes? Proteins In nature: Used by bacteria to cut viral DNA They “restrict” the growth of viruses http://www.dnalc.org/resources/animatio ns/restriction.htmlhttp://www.dnalc.org/resources/animatio ns/restriction.html

7. Restriction Enzymes Cut strands of DNA at specific nucleotide sequences There are many different restriction enzymes that each cut DNA at different nucleotide sequences Usually occurs at a palindrome 5’ GAATTC 3’ CTTAAG

8. What is a palindrome in English? Palindromes can be read backwards and forwards the same way. Give me an example of a name that is a palindrome: –Hannah It’s a little more tricky in DNA because the strand itself isn’t what must be backwards and forwards…

9. DNA palindromes A DNA palindrome reads exactly opposite its complementary strand Ex- CAGAC is not a palindrome but CCCGGG is. Why? Let’s look at the complementary strand: CAGACCCCGGG GUCUGGGGCCC

10. Another Example

11. Which of these pieces of DNA represents a palindrome? 1.5’ TTTCGC 3’ 3’ AAACCG 5’ 2.5’ GAATTC 3’ 5’ CTTAAG 3’ 3.5’ GAATTC 3’ 3’CTTAAG 5’ 4.5’AATGCC 3’ 5’ TTACGG 3’

12. Sticky Ends Most restriction enzymes cut DNA with a staggered cut The staggered cuts leave the DNA with end pieces “sticking off” –We call these “sticky ends” –These exposed N-bases will want to join with other complimentary exposed bases

13. So what ? What do you predict could happen if two pieces of DNA are cut with the same restriction enzyme??? –YES! They will have the same “sticky ends” –How could we use this???

14. R estriction E nzymes can be used to make R ecombinant DNA. The gene of interest can be isolated using R estriction E nzymes

15. Restriction Enzymes can be used to make RECOMBINANT DNA! The gene you are interested in inserting (aka the “gene of interest”) can be cut using a restriction enzyme. What will happen if I also cut the other organisms DNA with the same Restriciton Enzyme?

16. Making Recombinant DNA… Once the gene is isolated, how do we join it with the organism’s DNA? Cut the organism’s DNA with the same restriction enzyme…why –The sticky ends will naturally be attracted to each other Add DNA LIGASE: an enzyme that seals the fragments together

17. Applications of this technology PCR GMO’s- (Transgenic Organisms) Gene Therapy

18. PCR Now whenever the bacteria plasmid replicates, the gene will be cloned. Gene Cloning: the production of many identical copies of the same gene.

20. Transgenic OrganismYou have created a Transgenic Organism! –organisms that contain functional recombinant DNA (rDNA) from a different organism

21. Gene Therapy Cystic Fibrosis example http://learn.genetics.utah.edu/content/tech/ genetherapy/cysticfibrosis/http://learn.genetics.utah.edu/content/tech/ genetherapy/cysticfibrosis/

22. The Human Genome A map of all the base pairs in human DNA. Video- alopecia and the Human Genome Project