2 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 1Odds of blood on socks in bedroom not being N. Brown-Simpson’s:8.5 billion to 1Odds of blood on glove not being from R. Goldman, N. Brown-Simpson, and O.J. Simpson:21.5 billion to 1Number of people on planet earth:6.1 billionOdds of being struck by lightning in the U.S.:2.8 million to 1Odds of winning the Illinois Big Game lottery:76 million to 1Odds of getting killed driving to the gas station to buy a lottery ticket4.5 million to 1Odds of seeing 3 albino deer at the same time:85 million to 1Odds of having quintuplets:Odds of being struck by a meteorite:10 trillion to 1
3 Recombinant DNADef: DNA in which genes from 2 different sources are linkedGenetic engineering: direct manipulation of genes for practical purposesBiotechnology: manipulation of organisms or their components to perform practical tasks or provide useful products
5 DNA CloningRestriction enzymes (endonucleases): in nature, these enzymes protect bacteria from intruding DNA; they cut up the DNA (restriction); very specificRestriction site: recognition sequence for a particular restriction enzymeRestriction fragments: segments of DNA cut by restriction enzymes in a reproducable waySticky end: short extensions of restriction fragmentsDNA ligase: enzyme that can join the sticky ends of DNA fragmentsCloning vector: DNA molecule that can carry foreign DNA into a cell and replicate there (usually bacterial plasmids)
6 Steps for eukaryotic gene cloning Isolation of cloning vector (bacterial plasmid) & gene-source DNA (gene of interest)Insertion of gene-source DNA into the cloning vector using the same restriction enzyme; bind the fragmented DNA with DNA ligaseIntroduction of cloning vector into cells (transformation by bacterial cells)Cloning of cells (and foreign genes)Identification of cell clones carrying the gene of interest
7 DNA Analysis & Genomics PCR (polymerase chain reaction)Gel electrophoresisRestriction fragment analysis (RFLPs)Southern blottingDNA sequencingHuman genome project
8 Polymerase chain reaction (PCR) Amplification of any piece of DNA without cells (in vitro)Materials: heat, DNA polymerase, nucleotides, single-stranded DNA primersApplications: fossils, forensics, prenatal diagnosis, etc.
9 DNA AnalysisGel electrophoresis: separates nucleic acids or proteins on the basis of size or electrical charge creating DNA bands of the same length
10 Restriction fragment analysis Restriction fragment length polymorphisms (RFLPs)Southern blotting: process that reveals sequences and the RFLPs in a DNA sequenceDNA Fingerprinting
11 DNA SequencingDetermination of nucleotide sequences (Sanger method, sequencing machine)Genomics: the study of genomes based on DNA sequencesHuman Genome Project
12 Practical DNA Technology Uses Diagnosis of diseaseHuman gene therapyPharmaceutical products (vaccines)ForensicsAnimal husbandry (transgenic organisms)Genetic engineering in plantsEthical concerns?