1. Chapter 20 DNA Technology & Genomics

2. Slide 2 of 14 Biotechnology Terms Biotechnology Process of manipulating organisms or their components to make useful products Genetic engineering + tissue/cell culturing technologies Genetic Engineering Manipulation of genes and genome Insulin & GMO (Genetically Modified Organism) Recombinant DNA Artificially created DNA Typically, DNA is integrated from another species

3. Slide 3 of 14 Biotechnology Terms Gene Cloning Laboratory production of multiple copies of DNA segment Therapeutic cloning – embryonic stem cells Spinal cord injuries Reproductive (organismal) cloning – Dolly the sheep Restriction Enzymes Enzymes that cut DNA at specific locations Usually, derived from bacteria Cut sites of DNA = restriction fragments Sticky ends – restriction fragments usually have one end longer than the other

4. Slide 4 of 14

5. Slide 5 of 14

6. Slide 6 of 14 Cloning is fun!! 5 steps 1. Identify & isolate the gene of interest Involves finding a cloning vector – plasmid or organism used to carry the DNA sequence to be cloned 2. Restriction enzymes cut gene of interest from original site & cut vectors DNA This ensures matching sticky ends on gene of interest & vector DNA

7. Slide 7 of 14 Cloning is fun!! (Page 2) 5 steps 3. Form the recombinant plasmid Recombinant plasmid – plasmid + DNA fragments Sealed together using DNA Ligase Restriction enzymes cut gene of interest from original site & cut vectors DNA This ensures matching sticky ends on gene of interest & vector DNA 4. Get the vector (w/gene of interest) into a host cell Usually involves bacterial transformation 5. Select for cells that have been transformed Identify cells with gene of interest (antibiotic resistance)

8. Slide 8 of 14

9. Slide 9 of 14 PCR Polymerase Chain Reaction Thermal cycling Amplification of DNA 3 Steps Denaturation (Heating) Annealing (Cooling) Primer formation Extension DNA polymerase adds nucleotides at 3 end

10. Slide 10 of 14

11. Slide 11 of 14

12. Slide 12 of 14

13. Slide 13 of 14 Biotechnology Terms (More) Genomic library Thousands of recombinant plasmid clones Each has a piece of the original genome cDNA Library Complementary DNA Reverse transcriptase makes cDNA from mRNA

14. Slide 14 of 14