Cloning In Plasmids

Question 1

Step one

Answer 1

Grow human cells in culture (cells that contain gene of interest)

Question 2

Step two

Answer 2

Isolate the DNA from the cultured human cells. Cut the Gene X from the rest of the DNA, using a restriction enzyme to create sticky ends.

Question 3

Step three

Answer 3

Isolate a suitable plasmid vector from a bacterial such as E. coli

Question 4

Step four

Answer 4

Cut the plasmid with the same restriction enzyme as that used to cut Gene X. This produces a plasmid with matching sticky ends to the gene.

Question 5

Step five

Answer 5

The gene X is inserted into the plasmid by mixing the genre and the plasmids together. Some plasmids take up gene x. Three sticky ends of the plasmid base pair with the stocky ends of gene x.

Question 6

Step six

Answer 6

DNA ligase johns together the sticky ends of the plasmid and the gene, by covalent bonds

Question 7

Step seven

Answer 7

Three recombinant plasmid is now put back into the bacteria. The plasmids are added to a growing bacterial culture. Some bacteria will take up the plasmid by the process of “transformation”

Question 8

Step eight

Answer 8

The recombinant bacteria are now grown on an agar plate. Three bacteria multiply and reproduce many copies of the gene x. This genre has now been cloned. Colonies of bacteria containing the cloned gene are identified and the gene can be isolated from them for scientific or medical use.