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Biotechnology: Bacterial Transformation Lab

Terms in this set (25)

Is insulin usually made from engineered bacteria?
Yes
What allows molecular biologists to manipulate genetic information in a lab setting to understand more fully how DNA operates?
Plasmids !
What bacteria was transformed in this lab?
E. coli
What does the calculation of transformation efficiency tell us?
How well E. coli took up the foreign DNA
What are some key concepts that this lab focuses on?
Cell structure of bacteria; structure and function of cell membranes; enzymes; DNA; RNA; transcription; translation, evolution and interactions between organisms and their environment
Genetic variation can be introduced by what in bacteria?
Lateral transfer, transduction, or transformation
Many plasmids contain what?
Genes that code for resistance to antibiotics
Does swapping plasmids increase variation in bacteria?
Yes
When plasmids are inserted into an organism, what changes (besides the DNA sequence)?
The organism's phenotype
Plasmids are used as vectors. What does this mean?
Carriers of genetic information
Where in a human body can you find E. coli?
Colon
What is the gene for resistance to ampicillin called?
pAMP
Does this quizlet suck?
Hell ya
What is the transforming solution in the experiment?
CaCl2
Why are the E. coli cells "heat shocked" after they are transformed?
To keep the bacteria alive even after transformation
Why are the cells incubated in nutrient broth for a short time before being planted in agar?
To keep the bacteria alive even after transformation
On which plates would you expect to find bacteria most like the original non-transformed E. coli colonies you initially observed and why?
The plates with the -LB broth, + LB broth, and + LB/Amp both would be most like the original colonies because the bacteria would only be exposed to the nutrient broth and having the plasmid doesn't affect the amount of bacterial growth unless it is exposed to the ampR gene (Don't know if this is right, blame Mahima)
What evidence suggests that the changes were due to the transformation procedures that you performed?
The growth of bacteria in the + plasmid LB broth with ampicillin is evidence that the bacteria took in the plasmid containing the resistant gene. If they had not, they would have all died. Also, not having growth in the - plasmid plates is evidence that the plasmid affected bacterial growth.
What advantage would there be for an organism to be able to turn on or off particular genes in response to certain conditions?
If the bacteria is in the presence of ampicillin then it can turn on the resistant gene in order to combat death from ampicillin. However, if the bacteria isn't in presence of it, the gene can be turned off. This would help cells almost be able to control their own death (?)
Did the bacteria successfully take in the plasmid and how is this evident?
Yes, the bacteria that took in the plasmids had the most growth, which means the bacteria successfully took in the plasmid.
How do you calculate transformation efficiency?
# amp resistant colonies / ug of plasmid
What is the first step in calculating transformation efficiency?
Determine the mass of the plasmid added to the "+ plasmid" transformation reaction tube (concentration of plasmid x volume of plasmid added = ug plasmid in reaction).
What is the second step in calculating transformation efficiency?
Determine the concentration of plasmid in the reaction tube [ug plasmid / (broth + CaCl2) = concentration of plasmid in reaction]
What is the third step in calculating transformation efficiency?
Determine the mass of plasmid in the solution added to the agar plate (concentration of plasmid x volume of plasmid spread = ug plasmid on plate).
What is the fourth step in calculating transformation efficiency?
Determine the number of colonies on the "+ plasmid" LB?Amp plate per ug of plasmid (colonies of plasmid / ug of plasmid = transformation efficiency).
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