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23 Cards in this Set
- Front
- Back
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Bacterial Transformation?
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process where bacterial cells take up naked DNA molecules
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When will the DNA replicate the foreign DNA along with their own DNA?
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if the foreign DNA has an origin of replication recognized by the host cell DNA polymerases
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Competent?
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bacteria that are able to uptake DNA
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What are the 3 ways bacteria can be put into DNA?
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conjugation, transuction, transformation
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Conjugation?
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DNA is transferred from one bacterial donor cell to recipient cell via sex pilli
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Transduction?
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DNA is introduced into bacteria by injection from a virus
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Transformation?
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bacterial cells take up naked DNA molecules
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How does ampicillin prevent bacteria from growing?
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prevent bacteria from making a cell wall
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Why would you want to insert DNA into bacteria? 2
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-to make more DNA
-to make more protein |
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What will be inserted into E.Coli for this lab?
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pGLO plasmid
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What genes do pGLO contain?
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bla, GFP, araC
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Role of bla gene?
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provides resistance to the antibiotic ampicillin by using beta lactamase protein
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role of GFP gene?
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codes for green fluorescent protein
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Role of araC gene?
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promotes the production of GFP by producing araC protein
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How are competent bacteria made?
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treatment with calcium chloride followed by heat treatment at 42 degrees celsius
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Role of cold temperature in transformation?
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helps the DNA adhere to cell membrane of E.Coli
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Role of high temperatures in transformation?
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increases the permeability of the cell membrane to DNA
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What is used to help the bacteria recover from heat shock?
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briefly incubate bacteria with non-selective growith media (LB)
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Transformation efficiency?
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quantative value that describes how effective you were at getting a plasmid into bacteria
-represents the number of transformed colonies per microgram of DNA added |
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Transformation effiency formula?
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total number of cells growing on plate/amount of DNA spread on agar
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Formula to determine the amount of DNA spread on the agar(microg)?
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total amount of DNA(microg) *fraction of DNA used
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Formula to determine fraction of DNA used?
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volume spread on plate(micoL)/total sample volume in test tube (microL)
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Determine amount of pGLO in DNA(microg)?
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concentration of DNA (microg/microL) * volume of DNA (microL)
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