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 Prototroph Auxotroph Suppressor 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 4/19/2017

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 Genotype – Phenotypes Spontaneous Mutations Mutation Wild-Type Isolation Mutants Replica plating Auxotroph – Prototroph Penicillin-Selection Method Spontaneous Mutations Point Mutation Base-Pair Substitution Silent, Missense, Nonsense Mutations Mutation Genetic Recombination Microbial Models Molecular Cloning Biotechnology Mutants

Genetic Word List 2 Temperature Sensitive Mutation Reading Frame Frame Shift Mutation Back Mutations Suppressor Mutation Deletions Insertions Translocation Inversion Mutation Rate DNA ~10-6 RNA ~10-3 Mutagens Chemicals Base Analogs Intercalating Agents Radiation UV Light

Genetic Word List 3 Ionizing Radiation DNA Repair SOS Regulatory System Biological Mutagens Transposons Site Directed Mutations Carcinogenesis Ames Test Genetic Recombination Bacterial Gene Transfer Transformation Competent Transfection Electroporation Particle gun

Genetic Word List 4 Conjugation Plasmids Copy Number Incompatibility Curing Conjugative Plasmids Transduction Generalized Transduction Specialized Transduction Lysogeny Phage conversion Resistance Plasmids R Plasmids Virulence Plasmids Bacteriocin Engineered plasmids

Genetic Word List 5 Chromosomal Mobilization Sex Pili Fertility (F) Plasmids HFr Strains Interrupted Matings Genetic Maps Escherichia coli Chromosome 4,639,221 Base Pairs 4,288 Open Reading Frames Horizontal Gene Transfer Transposition Transposase Conservative – Replicative Transposition Phase Variation Inversion

Genetic Word List 6 Pathogenicity Islands Genomics Mycoplasma genitalium Genome 470 Open Reading Frames Genomic Mining