Gene Cloning

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  • Created by: Emaa.xo
  • Created on: 30-03-22 19:30
What is a vector?
A DNA molecule that is used to carry a foreign DNA into a host cell e.g. a bacterium cell.
Example of a vector is a plasmid (circular)
which will carry the genetic material.
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What properties do vectors need to have?
- It shouldn't already be a part of the host chromosome
- it should be able to be multiplied by the cell e.g. viruses in humans
- it should constantly be produced in the cell i.e. multiplication should be maintained
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How is the vector cut so that the gene of interest can be inserted into the vector (plasmid)?
Restriction endonuclease enzymes cut the vector at specific recognition sites.
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Why is it important for the restriction endonuclease enzymes to cut at just 1 specific recognition site?
Bc if the recognition site on the vector is not specific, the enzyme will cut open the entire vector, which means we cannot insert our gene of interest. The vector will just reform again, so 1 recognition site means the vector remains in shape.
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What is the role of DNA ligase in gene cloning?
They reseal the gaps in the phosphodiester backbone of the vector by attaching the gene through complementary base pairing.
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What is transformation in gene cloning?
The process of inserting the recombinant plasmid into a host cell
using CaCl2
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What is vector replication?
after the recombinant plasmid is inserted inside the host cell, it is the process when the vector produces multiple copies of the same vector
inside the host cell e.g. a bacterium.
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What is the selection / cell multiplication process?
The process when we specifically select for the host cells that contain the vector / the plasmid and multiply them.

We ignore the host cells without the plasmid bc they do not contain the gene we are interested in.
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What is secondary selection?
When we are able to mark the cell using colour to clearly identify the host cells containing the vector and gene of interest.
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What is the advantage of gene cloning?
We can use the multiple copies of the gene for:
1) expression profiling: e.g. genetically modified insulin to treat diabetes
2) investigation: to study the gene sequence and identify any potential targets for drugs.
3) Manipulation: allows for research
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Other cards in this set

Card 2

Front

What properties do vectors need to have?

Back

- It shouldn't already be a part of the host chromosome
- it should be able to be multiplied by the cell e.g. viruses in humans
- it should constantly be produced in the cell i.e. multiplication should be maintained

Card 3

Front

How is the vector cut so that the gene of interest can be inserted into the vector (plasmid)?

Back

Preview of the front of card 3

Card 4

Front

Why is it important for the restriction endonuclease enzymes to cut at just 1 specific recognition site?

Back

Preview of the front of card 4

Card 5

Front

What is the role of DNA ligase in gene cloning?

Back

Preview of the front of card 5
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