Chapter 20 DNA Technology & Genomics

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

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

Slide 4 of 14

Slide 5 of 14

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

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)

Slide 8 of 14

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

Slide 10 of 14

Slide 11 of 14

Slide 12 of 14

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

Slide 14 of 14