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Gene Transfer: How New Strains Arise and Biotechnology What special mechanisms allow bacteria to swap genes between cells? • Gene Transfer and Recombination • • Donor and recipient; recombination Prokaryotic Gene Transfer • Transformation • Transduction: Bacteriophage transfer • Conjugation • Plasmids: F factors, R factors • Eukaryotic Gene Recombination • Viral Gene Changes • Application of DNA Biotechnology • • Fast, inexpensive, ethical methods Strategies of Gene Manipulation/Analysis • Recombinant DNA and Cloning • Restriction enzymes • Steps of Cloning the Insulin Gene • Making many Copies: Polymerase Chain Reaction • DNA Fingerprinting • Genetic sequencing Recombination Produces New Genotypes Donor DNA Recipient DNA Genotype: abcde Genotype: aBcde (now has a new phenotype or ability, like being able to use a sugar it could not before) Figure 8.25 Transformation: Absorption of Free DNA In a lab setting, many bacterial species are not “competent” to take up donor DNA. They are incubated in CaCl2 to make them competent and then heat shocked so they will suck up DNA fragments from the medium. Figure 8.24 Plasmids: Autonomously Replicating Extrachromosomal DNA Fertility (F) factor is a plasmid that can direct the formation of a sex pilus and DNA transfer by conjugation Conjugation: Simple F+ to F- Transfer Figure 8.27a Conjugation: Creation of an Hfr Cell Figure 8.27b Conjugation: Hfr to F- cell Figure 8.27c Transduction Phage protein coat Bacterial chromosome Recombinant Bacterial DNA Donor bacterial DNA Phage DNA Recipient cell Recipient bacterial DNA Recombinant cell Figure 8.28 Gene Transfer: How New Strains Arise and Biotechnology What special mechanisms allow bacteria to swap genes between cells? • Gene Transfer and Recombination • Donor and recipient; recombination • Prokaryotic Gene Transfer • Transformation • Transduction: Bacteriophage transfer • Conjugation • Plasmids: F factors, R factors • Eukaryotic Gene Recombination • Viral Gene Changes • Application of DNA Biotechnology • Fast, inexpensive, ethical methods • Strategies of Gene Manipulation/Analysis • Recombinant DNA and Cloning • Restriction enzymes • Steps of Cloning the Insulin Gene • Making many Copies: Polymerase Chain Reaction • DNA Fingerprinting Genetic Recombination In Eukaryotes: Crossing Over • Prophase I of Meiosis Figure 8.23 Double Infection By Two Viruses Can Produce a New Novel Virus By Recombination (In eukaryotes, is this how new recombinations of influenza viruses arise) Gene Transfer: How New Strains Arise and Biotechnology What special mechanisms allow bacteria to swap genes between cells? • Gene Transfer and Recombination • Donor and recipient; recombination • Prokaryotic Gene Transfer • Transformation • Transduction: Bacteriophage transfer • Conjugation • Plasmids: F factors, R factors • Eukaryotic Gene Recombination • Viral Gene Changes • Application of DNA Biotechnology • Fast, inexpensive, ethical methods • Strategies of Gene Manipulation/Analysis • Recombinant DNA and Cloning • Restriction enzymes • Steps of Cloning the Insulin Gene • Making many Copies: Polymerase Chain Reaction • DNA Fingerprinting Biotechnology/DNA Technology: Genetic Engineering Scheme Advantages & Disadvantages Figure 9.1.1 Restriction Enzymes Recognize and Cut at Recognition Sites Figure 9.2 Vector: Something That Can Carry in New DNA and Be Stable in the Cell Should have several restriction enzyme sites Figure 9.3 Gene Transfer: How New Strains Arise and Biotechnology What special mechanisms allow bacteria to swap genes between cells? • Gene Transfer and Recombination • Donor and recipient; recombination • Prokaryotic Gene Transfer • Transformation • Transduction: Bacteriophage transfer • Conjugation • Plasmids: F factors, R factors • Eukaryotic Gene Recombination • Viral Gene Changes • Application of DNA Biotechnology • Fast, inexpensive, ethical methods • Strategies of Gene Manipulation/Analysis • Recombinant DNA and Cloning • Restriction enzymes • Steps of Cloning the Insulin Gene • Making many Copies: Polymerase Chain Reaction • DNA Fingerprinting Genetic Engineering Involves Insertion of a Gene on a Plasmid Safety Issues and Ethics • Avoid accidental release of supergerms • Genetically modified crops must be safe for consumption and for the environment • Who will have access to an individual's genetic information? • Do we have a right to move genes around, creating new life forms (i.e. “playing God”)? Gene Transfer: How New Strains Arise and Biotechnology What special mechanisms allow bacteria to swap genes between cells? • Gene Transfer and Recombination • • Donor and recipient; recombination Prokaryotic Gene Transfer • Transformation • Transduction: Bacteriophage transfer • Conjugation • Plasmids: F factors, R factors • Eukaryotic Gene Recombination • Viral Gene Changes • Application of DNA Biotechnology • • Fast, inexpensive, ethical methods Strategies of Gene Manipulation/Analysis • Recombinant DNA and Cloning • Restriction enzymes • Steps of Cloning the Insulin Gene • Making many Copies: Polymerase Chain Reaction • DNA Fingerprinting • Genetic sequencing Polymerase Chain Reaction: DNA Replication in a Test Tube Exponential Increase in the Number of DNA Molecules each Cycle Gene Transfer: How New Strains Arise and Biotechnology What special mechanisms allow bacteria to swap genes between cells? • Gene Transfer and Recombination • • Donor and recipient; recombination Prokaryotic Gene Transfer • Transformation • Transduction: Bacteriophage transfer • Conjugation • Plasmids: F factors, R factors • Eukaryotic Gene Recombination • Viral Gene Changes • Application of DNA Biotechnology • • Fast, inexpensive, ethical methods Strategies of Gene Manipulation/Analysis • Recombinant DNA and Cloning • Restriction enzymes • Steps of Cloning the Insulin Gene • Making many Copies: Polymerase Chain Reaction • DNA Fingerprinting • Genetic Sequencing DNA Fingerprinting Generates Banding Patterns Unique to Individuals 1. Collect cells 2. Extract DNA 3. Cut the DNA in fragments using the same restriction enzyme 4. Separate the fragments using gel electrophoresis Figure 12.12 Gene Transfer: How New Strains Arise and Biotechnology What special mechanisms allow bacteria to swap genes between cells? • Gene Transfer and Recombination • • Donor and recipient; recombination Prokaryotic Gene Transfer • Transformation • Transduction: Bacteriophage transfer • Conjugation • Plasmids: F factors, R factors • Eukaryotic Gene Recombination • Viral Gene Changes • Application of DNA Biotechnology • • Fast, inexpensive, ethical methods Strategies of Gene Manipulation/Analysis • Recombinant DNA and Cloning • Restriction enzymes • Steps of Cloning the Insulin Gene • Making many Copies: Polymerase Chain Reaction • DNA Fingerprinting • Genetic sequencing Genetic Sequencing • Comparing the nucleotide sequences of rRNA genes(the 16S type) in prokaryotes (and eukaryotes) has helped show evolutionary relatedness • Used to separate prokaryotes and eukaryotes into 3 Domains in the late 1980s Gene Transfer: How New Strains Arise and Biotechnology What special mechanisms allow bacteria to swap genes between cells? • Gene Transfer and Recombination • • Donor and recipient; recombination Prokaryotic Gene Transfer • Transformation • Transduction: Bacteriophage transfer • Conjugation • Plasmids: F factors, R factors • Eukaryotic Gene Recombination • Viral Gene Changes • Application of DNA Biotechnology • • Fast, inexpensive, ethical methods Strategies of Gene Manipulation/Analysis • Recombinant DNA and Cloning • Restriction enzymes • Steps of Cloning the Insulin Gene • Making many Copies: Polymerase Chain Reaction • DNA Fingerprinting • Genetic sequencing