Ch. 20 - Biotechnology “Trial of the Century”:1994 Murder Case: Nicole Brown Simpson Blood Trail (1:40…) DNA Fingerprinting Animation

O.J. Simpson capital murder case,1/95-9/95 Odds of blood in Ford Bronco not being R. Goldman’s: 6.5 billion to 1 Odds of blood on socks in bedroom not being N. Brown-Simpson’s: 8.5 billion to 1 Odds of blood on glove not being from R. Goldman, N. Brown-Simpson, and O.J. Simpson: 21.5 billion to 1 Number of people on planet earth: 7 billion Odds of being struck by lightning in the U.S.: 2.8 million to 1 Odds of seeing 3 albino deer at the same time: 85 million to 1 DID HE DO IT??

DNA Analysis Gel electrophoresis: separates nucleic acids or proteins on the basis of size or electrical charge creating DNA bands of the same length Electrophoresis lab: DNA size vs. migration of DNA fragment tutorial

DNA Profiling VNTR & STR (“stutters”) – Variable Nucleotide Tandem Repeats Short Tandem Repeats. PCR – uses thermo-cycler & Taq polymerase to amplify amounts of DNA RFLPs - Restriction fragment length polymorphisms, generated from differences in VNTR’s & STR’s. Southern blotting: process that reveals sequences and the RFLPs in a DNA sequence. Animation

FBI & Combined DNA Index System

VOCAB Recombinant DNA: DNA in which genes from 2 different sources are combined Genetic engineering: direct manipulation of genes for practical purposes Biotechnology: manipulation of organisms or their components to perform practical tasks or provide useful products

Restriction Map of Lambda DNA

DNA Cloning Restriction enzymes (endonucleases): in nature, these enzymes protect bacteria from intruding DNA; they cut up the DNA (restriction); very specific Restriction site: recognition sequence for a particular restriction enzyme Restriction fragments: segments of DNA cut by restriction enzymes in a reproducable way Sticky end: short extensions of restriction fragments DNA ligase: enzyme that can join the sticky ends of DNA fragments Cloning vector: DNA molecule that can carry foreign DNA into a cell and replicate there (usually bacterial plasmids)

Reproductive Cloning Therapeutic cloning treats disease via “replacement cells”, tissues Reproductive cloning copies entire animal Uses? Ethics? Activity: What is Cloning?

Stem Cells Embryonic stem cells (PS or iPS) vs. adult stem cells

Can stem cells mend a broken heart? Unlocking Potential of Stem Cells

Practical Use of DNA Technology MEDICAL Diagnosis, treatment, prevention of genetic & infectious diseases Human gene therapy: Cystic Fibrosis Trials Abnormal allele Pharmaceuticals: proteins as drugs via gen engineered cells or ‘pharm’ animals (goats that make antithrombin) DNA Microarrays (gene chips) that monitor gene expression Forensics & Genetic Profiling DNA fingerprinting to exclude/identify criminal suspects, paternity Environment – oil cleanup via bacteria Agriculture & GM foods “Frankenfood” - Ethics Activity DNA microarray animation Microarray Virtual Lab

Plasmids, gene cloning, applications Animation

Transformation Lab: Engineering antibiotic resistance in bacteria Use plasmid as vector & E. coli colony in log phase growth CaCl2 + heat shock to induce plasmid uptake by bacteria (make cells “competent”) Grow on antibiotic medium to select for transformed cells