Lecture

1
Lecture 9 Date _____

• Chapter 20 DNA Technology Genomics

2
O.J. Simpson capital murder case,1/95-9/95

• Odds of blood in Ford Bronco not being R.
  Goldmans
• 6.5 billion to 1
• Odds of blood on socks in bedroom not being N.
  Brown-Simpsons
• 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
• 6.1 billion
• Odds of being struck by lightning in the U.S.
• 2.8 million to 1
• Odds of winning the Illinois Big Game lottery
• 76 million to 1
• Odds of getting killed driving to the gas station
  to buy a lottery ticket
• 4.5 million to 1
• Odds of seeing 3 albino deer at the same time
• 85 million to 1
• Odds of having quintuplets
• 85 million to 1
• Odds of being struck by a meteorite

3
Recombinant DNA

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

4
Bacterial plasmids in gene cloning
5
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)

6
Restriction Enzymes
7
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 ligase
• Introduction of cloning vector into cells
  (transformation by bacterial cells)
• Cloning of cells (and foreign genes)
• Identification of cell clones carrying the gene
  of interest

8
Cloning
9
DNA Analysis Genomics

• PCR (polymerase chain reaction)
• Gel electrophoresis
• Restriction fragment analysis (RFLPs)
• Southern blotting
• DNA sequencing
• Human genome project

10
Polymerase chain reaction (PCR)

• Amplification of any piece of DNA without cells
  (in vitro)
• Materials heat, DNA polymerase, nucleotides,
  single-stranded DNA primers
• Applications fossils, forensics, prenatal
  diagnosis, etc.

11
DNA Analysis

• Gel electrophoresis separates nucleic acids or
  proteins on the basis of size or electrical
  charge creating DNA bands of the same length

12
Restriction fragment analysis

• Restriction fragment length polymorphisms (RFLPs)
• Southern blotting process that reveals
  sequences and the RFLPs in a DNA sequence
• DNA Fingerprinting

13
DNA Sequencing

• Determination of nucleotide sequences (Sanger
  method, sequencing machine)
• Genomics the study of genomes based on DNA
  sequences
• Human Genome Project

14
Practical DNA Technology Uses

• Diagnosis of disease
• Human gene therapy
• Pharmaceutical products (vaccines)
• Forensics
• Animal husbandry (transgenic organisms)
• Genetic engineering in plants
• Ethical concerns?