UCSC BIOLOGY 119 - Mutations by Adding New DNA to the Genome

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I. Mutations by adding new DNA to the genomeA. DNA can be taken up by prokaryotes and incorporated into their genomesB. DNA that comes in either can replicate on its own, or is recombined into the chromosome1. To replicate on its own:2. Otherwise, to persist it has to go into:3. How this is done (Figure 15.6 and 15.7):C. Getting DNA into cell by transformation:1. Taking up plain old DNA2. Natural Competence: story of Streptococcus pneumoniae (Fig 15.14)3. Artificially induced competenceD. Getting DNA into a cell by Conjugation1. Transfer of genes from one prokaryote to another by a mechanism involving cell-to-cell contact2. What is transferred?3. How conjugation works (Figure 15.9):4. How scientists figured out cell-to-cell contact was required for conjugation. (Box 15.1 U tube experiment)E. Getting DNA by Transduction:F. Using bacterial mutagenesis1. Common type: gene replacement in bacteria (Fig. 15.11 and 15.12)a) Method for adding or deleting a geneb) Why do this?G. Random transposon mutagenesis (Fig. 15.17 and 15.18)1. What is a transposon?a) An genetic element that moves from one site into anotherb) The Tn is has two inverted repeatsc) Usually has some antibiotic resistance marker in between IRe) The Tn is also engineered without the transposase so that it won’t transpose again.2. Screen for mutants (takes longer to do)3. Selection for mutants (harder to do with for Tn mutagenesis)Mutations Part II Bio119I. Mutations by adding new DNA to the genomeA. DNA can be taken up by prokaryotes and incorporated into their genomesB. DNA that comes in either can replicate on its own, or is recombined into the chromosome1. To replicate on its own:2. Otherwise, to persist it has to go into:3. How this is done (Figure 15.6 and 15.7):C. Getting DNA into cell by transformation:1. Taking up plain old DNA2. Natural Competence: story of Streptococcus pneumoniae (Fig 15.14)Page 1Mutations Part II Bio1193. Artificially induced competenceD. Getting DNA into a cell by Conjugation1. Transfer of genes from one prokaryote to another by a mechanism involving cell-to-cell contact2. What is transferred? 3. How conjugation works (Figure 15.9):4. How scientists figured out cell-to-cell contact was required for conjugation. (Box 15.1 U tube experiment)E. Getting DNA by Transduction:F. Using bacterial mutagenesis1. Common type: gene replacement in bacteria (Fig. 15.11 and 15.12)Page 2Mutations Part II Bio119a) Method for adding or deleting a geneb) Why do this?G. Random transposon mutagenesis (Fig. 15.17 and 15.18)1. What is a transposon?a) An genetic element that moves from one site into anotherb) The Tn is has two inverted repeatsc) Usually has some antibiotic resistance marker in between IRd)e) The Tn is also engineered without the transposase so that it won’t transpose again.Page 3Mutations Part II Bio119f)2. Screen for mutants (takes longer to do)3. Selection for mutants (harder to do with for Tn mutagenesis)Page


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