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Microbial Genetics

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.

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Microbial Genetics

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  1. Microbial Genetics • Mutation • Genetic Recombination • Model organism • E. coli • Molecular Cloning • Bioinformatics “Golden” Rice

  2. Plasmid Types • Resistance Plasmids – R Plasmids • Virulence Plasmids • Bacteriocins • Engineered Plasmids

  3. Mutants • Mutation • Wild Type • Genotype • Phenotype

  4. Isolation of Mutants • Replica Plating • Auxotrophs – Nutritional Mutants • Prototroph – Wild Type

  5. Penicillin Selection Method – Kills Growing Cells

  6. Mutations • Spontaneous Mutations • Point Mutations • Base-Pair Substitution • Silent, • Missense, or • Nonsense Mutations

  7. Conditional Lethal Mutations • Temperature Sensitive Mutations Incubation Temperature 30 C 40 C

  8. Reading Frame Frame Shift Mutation

  9. Reversion Mutation Prototroph Auxotroph Back Mutation • Back • Mutation • Suppressor • Mutation Mutation Back Mutation Normal Protein

  10. 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

  11. Inversion Orientation of a Piece of DNA is Reversed in the Genome - Phase Variation

  12. 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

  13. Mutagens • Chemicals • Base Analogs – Chemically Resemble Base

  14. Intercalating Agent • Inserts Between Base Pairs

  15. Radiation • UV Light • Ionizing Radiation • DNA Repair • SOS Regulatory System

  16. Biological Mutagens • Transposons – Can Insert Into Gene • Transposon Mutagenesis • Site Directed Mutations – Man-made

  17. Ames Test • Mutagens  Carcinogens • Ames Test Measures Back Mutation Rates to Detect Carcinogens • Liver extract

  18. Bacterial Gene Transfer • Genetic Recombination • 3 Types of Bacterial Gene Exchange: • 1. Transformation • “Naked DNA” • Competent

  19. Transfection in Eukaryotes • Particle (Gene) Guns Insert DNA • Electroporation

  20. 2. Transduction • Generalized Transduction

  21. Specialized Transduction

  22. Lysogeny Can Lead to Phage Conversion

  23. 3. Conjugation • Plasmids • Copy Number • Incompatibility • Curing • Conjugative Plasmid

  24. Chromosomal Mobilization Fertility (F) Plasmid Episome HFr Strain Sex Pili

  25. Gene Maps • Interrupted Matings

  26. E. coli Chromosome • 4,639,221 Base Pairs • 4,288 Open Reading Frames • 88% ORF • 1% rRNA, tRNA • 10% Promoter, Origin, etc • 6% Operons

  27. Lateral Gene Transfer • Horizontal Gene transfer • Pathogenicity Islands • ICE – Integrative and Conjugative Elements

  28. Transposition • Transposase • Conservative Transposition • Replicative Transposition • Phase Variation • Inversion

  29. Genomics • Mycoplasma genitalium Genome • 470 Open Reading Frame • Genomic Mining

  30. Genetic Word List Genotype – Phenotypes 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

  31. 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

  32. 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

  33. 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

  34. 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

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

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