Microbial Engineering
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| Fig. 9.2 | Restriction Enzymes (2 Figures)
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| Fig. 9.4 | PCR (2 Figures)
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| Fig. 9.16 | Southern Blotting
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| What are the steps of generalized transduction? FIRST STEP ONLY PLEASE | Bacteriophage attaches to cell wall and injects its DNA to bacteria
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| What is the second step of transduction? | Phage DNA and proteins are made from DNA released and bacterial chromosome is broken into pieces by phage enzyme
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| THIRD POSSIBLE STEP? | (Bacterial DNA gets into phage capsid), then the donor cell lyses and relases phages
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| STEP 4 | Phage infects new host cell
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| Step 5 | REcombination can occur, can produce recombinant cell with a genotype different from botth donor and recipient
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| Why would specialized transduction occur? | When phage wants something specific from the bacteria, such as a toxin
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| What is the core difference between the specialized vs. generalized transduction? | The fact that a specific DNA region is transducted.
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| GENETIC ENGINEERING | GENETIC ENGINEERING
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| Thus, what are two key tools in genetic engineering? | Restriction enzymes and cloning plasmids
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| What are restriction enzymes? | DNA cutting enzymes in bacteria that recognizes and cuts only one particular sequence of nucleotide bases in DNA and it cuts them the same way each time
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| What is the process of restriction enzyme in making rDNA? What is needed to form rDNA? | The restriction enzyme cuts the double stranded DNA at a speciific site -->sticky ends-->identical sets join by base pairing --> make rDNA by DNA ligase
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| PCR | PCR
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| What kind of enzyme is used to copy DNA in PCR? What kind of bcteria is needed? | DNA polymerase from thermophillic bacteria that can survive extreme 94 degree heat
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| What is the process of PCR? | 1) Get DNA, 2) Melt it w/ heat stable polymerase 3) Add primers 4) Add heat to melt DNA strand together 5) Repeat
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| What are primers? Why do we need them? | Sequences of DNA specfic to gene that will indicate WHAT we will be copying
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| What is PCR? | Making a lot of DNA from little starting material
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| What do we do when we get the gene that we want? Where do we put them? | Bacterial fingerprinting to see if bacteria are related, Gene therapy
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| Is Gene therapy a successful idea? | Nope...
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| DNA probes? | Short segments of single stranded DNA that are complementary to the desired gene
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| What is southern blotting? | Detection of DNA on filter q/ restriction enzymes without much radioactive probes
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| What has replaced the process of BActerial DNA fingerprinting? | PCR reaction
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| Introns vs. extrons and cDNA | Figure 9.9
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| How do you make cDNA? | 1. RNA polymerase transcribes DNA w/ introns and extrons-->RNA 2. Remove introns 3. Isolate mRNA (add RT) 4. Make first DNA 5. Make second 6. RT digests mRNA (making cDNA w/ no introns)
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