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Microbial Genetics • Mutation • Genetic Recombination • Model organism – E. coli • Molecular Cloning • Bioinformatics “Golden” Rice Plasmid Types • Resistance Plasmids – R Plasmids • Virulence Plasmids • Bacteriocins • Engineered Plasmids Mutants • • • • Mutation Wild Type Genotype Phenotype Isolation of Mutants • Replica Plating • Auxotrophs – Nutritional Mutants • Prototroph – Wild Type • Penicillin Selection Method – Kills Growing Cells Mutations • Spontaneous Mutations • Point Mutations • Base-Pair Substitution • Silent, • Missense, or • Nonsense Mutations Conditional Lethal Mutations • Temperature Sensitive Mutations Incubation Temperature 30 C 40 C Reading Frame Frame Shift Mutation Reversion Mutation • Back Mutation • Suppressor Prototroph Auxotroph Mutation Back Mutation Mutation Back Mutation Normal Protein Other Mutations • Deletions – Lose of a Piece of DNA • Insertions – Additions of a Piece of DNA • Translocation – Movement of a Piece of DNA to a New Location in the Genome Inversion Orientation of a Piece of DNA is Reversed in the Genome - Phase Variation Mutation Rate • Spontaneous (Natural or Background) Mutation Rate • DNA ~ 1 per Million (varies between genes and location) • RNA ~ 1 per Thousand • Mutagens – Anything that Increases the Mutation Rate Mutagens • Chemicals • Base Analogs – Chemically Resemble Base • Intercalating Agent Inserts Between Base Pairs • Radiation • UV Light • Ionizing Radiation • DNA Repair • SOS Regulatory System 5/23/2017 15 Biological Mutagens • Transposons – Can Insert Into Gene • Transposon Mutagenesis • Site Directed Mutations – Man-made Ames Test • Mutagens Carcinogens • Ames Test Measures Back Mutation Rates to Detect Carcinogens • Liver extract Bacterial Gene Transfer • Genetic Recombination • 3 Types of Bacterial Gene Exchange: • 1. Transformation • “Naked DNA” • Competent • Transfection in Eukaryotes • Particle (Gene) Guns Insert DNA •Electroporation 2. Transduction • Generalized Transduction • Specialized Transduction • Lysogeny Can Lead to Phage Conversion 3. Conjugation • • • • • Plasmids Copy Number Incompatibility Curing Conjugative Plasmid Chromosomal Mobilization • • • • Fertility (F) Plasmid Episome HFr Strain Sex Pili Gene Maps • Interrupted Matings E. coli Chromosome • 4,639,221 Base Pairs • 4,288 Open Reading Frames • 88% ORF • 1% rRNA, tRNA • 10% Promoter, Origin, etc • 6% Operons Lateral Gene Transfer • Horizontal Gene transfer • Pathogenicity Islands • ICE – Integrative and Conjugative Elements Transposition • Transposase • Conservative Transposition • Replicative Transposition • Phase Variation • Inversion Genomics • Mycoplasma genitalium Genome • 470 Open Reading Frame • Genomic Mining Genetic Word List Mutation Genotype – Phenotypes Spontaneous Mutations Genetic Recombination Wild-Type Point Mutation Isolation Mutants Base-Pair Substitution Microbial Models Molecular Cloning Biotechnology Mutants Replica plating Auxotroph – Prototroph Silent, Missense, Nonsense PenicillinSelection Method Mutations Genetic Word List 2 Temperature Sensitive Mutation Reading Frame Frame Shift Mutation Back Mutations Suppressor Mutation Deletions Mutagens Insertions Chemicals Translocation Base Analogs Inversion Intercalating Agents Mutation Rate DNA ~10-6 RNA ~10-3 Radiation UV Light Genetic Word List 3 Ionizing Radiation DNA Repair Site Directed Mutations Bacterial Gene Transfer SOS Regulatory System Carcinogenesi s Transformation Biological Mutagens Ames Test Transposons Genetic Recombination Competent Transfection Electroporation Particle gun Genetic Word List 4 Transduction Conjugation Generalized Transduction Plasmids Resistance Plasmids Copy Number R Plasmids Specialized Transduction Incompatibility Virulence Plasmids Lysogeny Phage conversion Curing Bacteriocin Conjugative Plasmids Engineered plasmids Genetic Word List 5 Chromosomal Mobilization Sex Pili Fertility (F) Plasmids HFr Strains Interrupted Matings Genetic Maps Transposition Transposase Escherichia coli Chromosome 4,639,221 Base Conservative – Pairs Replicative 4,288 Open Transposition Reading Frames Phase Horizontal Gene Variation Transfer Inversion Genetic Word List 6 Pathogenicity Islands Genomics Mycoplasma genitalium Genome 470 Open Reading Frames Genomic Mining
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