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MICRO2

Microbial Engineering

QuestionAnswer
Fig. 9.2 Restriction Enzymes (2 Figures)
Fig. 9.4 PCR (2 Figures)
Fig. 9.16 Southern Blotting
What are the steps of generalized transduction? FIRST STEP ONLY PLEASE Bacteriophage attaches to cell wall and injects its DNA to bacteria
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
THIRD POSSIBLE STEP? (Bacterial DNA gets into phage capsid), then the donor cell lyses and relases phages
STEP 4 Phage infects new host cell
Step 5 REcombination can occur, can produce recombinant cell with a genotype different from botth donor and recipient
Why would specialized transduction occur? When phage wants something specific from the bacteria, such as a toxin
What is the core difference between the specialized vs. generalized transduction? The fact that a specific DNA region is transducted.
GENETIC ENGINEERING GENETIC ENGINEERING
Thus, what are two key tools in genetic engineering? Restriction enzymes and cloning plasmids
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
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
PCR PCR
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
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
What are primers? Why do we need them? Sequences of DNA specfic to gene that will indicate WHAT we will be copying
What is PCR? Making a lot of DNA from little starting material
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
Is Gene therapy a successful idea? Nope...
DNA probes? Short segments of single stranded DNA that are complementary to the desired gene
What is southern blotting? Detection of DNA on filter q/ restriction enzymes without much radioactive probes
What has replaced the process of BActerial DNA fingerprinting? PCR reaction
Introns vs. extrons and cDNA Figure 9.9
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)
Created by: talkglitter2486
 

 



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