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What type of gene is used to distinguish bacteria that carry a plasmid containing foreign DNA from those that don' t?
The plasmid that contains foreign DNA is engineered to also carry an antibiotic resistance gene. This antibiotic resistance gene codes for a protein that is able to inactivate an antibiotic thus keeping the cell alive. In the absence of the antibiotic resistance gene, the cells would not survive when exposed to an antibiotic. After transfection (the process of inserting the plasmid carrying the foreign gene into cells), the cells are gown in media containing an antibiotic. Cells that contain the plasmid (and therefore contain the antibiotic resistance gene) are able to survive in this medium. Cells that do not contain the plasmid (and therefore lack the antibiotic resistance gene) do not survive in this medium. The process described above is called selection
How will you identify cells carrying the recombinant plasmid?
The transformants are selected for on agar containing an appropriate antibiotic. For example if your recombinant plasmid contains a kanamycin cassette, then only the cells containing the plasmid will grow on an agar plate containing kanamycin. PCR can then be performed on the colonies to ensure they contain your gene of interest on the plasmid.
Can transformation only be performed using plasmid DNA containing antibiotic resistant genes R factors?
No, transformation can occur using a variety of methods and does not require the presence of antibiotic resistant genes. While antibiotic resistance can be a useful marker in selection processes, it is not a necessary component for transformation to take place.
What type of plasmid is the source of the problem with antibiotic resistance?
The source of antibiotic resistance is often plasmids known as R plasmids, which carry genes that confer resistance to antibiotics. These plasmids can be transferred between bacteria, spreading antibiotic resistance throughout microbial populations.
Why is ampicillin added to the pound medium?
Ampicillin is an antibiotic that is usually used as a reporter gene in cloning. A plasmid containing the ampicillin resistance gene (as well as another target gene within the plasmid) is introduced into the bacterial host. If the bacterium has taken up the plasmid and is expressing the plasmid, it will be resistant to ampicillin. LB is used as a growth medium and ampicillin to verify the plasmid is within the bactrium. No growth means no plasmid in the bacterial host...
Is it true cells containing a plasmid with an antibiotic resistance gene will die in the presence of the antibiotic?
false
What type of gene is used to distinguish bacteria that carry a plasmid containing foreign DNA from those that don' t?
The plasmid that contains foreign DNA is engineered to also carry an antibiotic resistance gene. This antibiotic resistance gene codes for a protein that is able to inactivate an antibiotic thus keeping the cell alive. In the absence of the antibiotic resistance gene, the cells would not survive when exposed to an antibiotic. After transfection (the process of inserting the plasmid carrying the foreign gene into cells), the cells are gown in media containing an antibiotic. Cells that contain the plasmid (and therefore contain the antibiotic resistance gene) are able to survive in this medium. Cells that do not contain the plasmid (and therefore lack the antibiotic resistance gene) do not survive in this medium. The process described above is called selection
How will you identify cells carrying the recombinant plasmid?
The transformants are selected for on agar containing an appropriate antibiotic. For example if your recombinant plasmid contains a kanamycin cassette, then only the cells containing the plasmid will grow on an agar plate containing kanamycin. PCR can then be performed on the colonies to ensure they contain your gene of interest on the plasmid.
Can transformation only be performed using plasmid DNA containing antibiotic resistant genes R factors?
No, transformation can occur using a variety of methods and does not require the presence of antibiotic resistant genes. While antibiotic resistance can be a useful marker in selection processes, it is not a necessary component for transformation to take place.
What structures contains genes for enzymes and antibiotic resistance?
plasmid
What type of plasmid is the source of the problem with antibiotic resistance?
The source of antibiotic resistance is often plasmids known as R plasmids, which carry genes that confer resistance to antibiotics. These plasmids can be transferred between bacteria, spreading antibiotic resistance throughout microbial populations.
Why is ampicillin added to the pound medium?
Ampicillin is an antibiotic that is usually used as a reporter gene in cloning. A plasmid containing the ampicillin resistance gene (as well as another target gene within the plasmid) is introduced into the bacterial host. If the bacterium has taken up the plasmid and is expressing the plasmid, it will be resistant to ampicillin. LB is used as a growth medium and ampicillin to verify the plasmid is within the bactrium. No growth means no plasmid in the bacterial host...
What is an extra loop of DNA that contains antibiotic resistance what gene?
An extra loop of DNA that carries antibiotic resistance genes is called a plasmid. These genes can provide bacteria with the ability to survive exposure to antibiotics.
What one plate would you first inspect to conclude that the transformation occurred successfully Why?
If you transform bacteria with a plasmid containing a selection marker (such as an antibiotic resistance gene) and plate the transformed bacteria on a plate suited for selecting for plasmid-containing bacteria (such as a plate containing an antibiotic that only those bacteria with antibiotic resistance can survive), then simply inspecting whether colonies are present on the plate will suffice in determining whether the transformation succeeded. If no colonies are found, that means no bacteria got the antibiotic resistance gene on the plasmid and the transformation was unsuccessful. If some colonies are found, that means some bacteria contain the plamis containing the antibiotic resistance gene and those colonies can the transformation was successful.
What is one way to determine whether a bacterial culture has received a recombinant plasmid?
The plasmid have a "reporter gene" inside it, generally resistance to specific antibiotic. the plasmid is transformed into bacteria that don't have resistance to that specific antibiotic drug, and than the cultured on a petri-dish that contain the antibiotic drug. Only bacteria that had receive the plasmid will have resistance and grow, all the other will die.
How does plasmid allow for antibiotic resistance?
A plasmid (conjugative plasmid) that has a resistance gene on it, can transfer itself to another bacterial cell (called conjugation) or assist in the transfer of a non-conjugative plasmid that has a resistance gene to another cell (called mobilization). Whichever way it happens, once the plasmid is transfered to the new cell, this cell too may show signs of resistance to the particular antibiotic. This is one of the ways of the speard of resistance amongst bacteria
You have taken two control.One with LB media only and other with LB plus kanamycin.LB plus kanamycin showed growth upon incubation. what could be the reason?
This means the bacteria that you are handling is resistant to kanamycin and can therefore grow in the presence of this antibiotic. You have to check if there are colonies on the control plate containing only LB medium. If there are colonies, then this reasoning is valid. A possible reason why bacteria can grow in the presence of kanamycin is they could have a plasmid that is conferring antibiotic resistance. However, this is only a hypothesis and further investigation (like plasmid isolation) is required to confirm it.
