B6 Gene Technologies

Question 1

what is genetic engineering

Answer 1

a technique used to transfer genes from one organism to another

Question 2

before genetic engineering occurs, what must we do first

Answer 2

1. Isolate the desired

gene.

Question 3

what is the first step of genetic engineering and what does this produce

Answer 3

Cut the gene using restriction enzymes. The enzyme cuts the restriction site at the two ends of the gene to produce ‘sticky ends’ (single-stranded DNA).

Question 4

once cutting the gene of interest with restriction enzymes, what do we do next

Answer 4

Insert the gene into the vector

DNA (the DNA of another organism such as a bacterium or a virus).

Question 5

how do we prepare the vector to insert the gene of interest that has been cut by restriction enzymes

Answer 5

The same restriction enzymes that were used to cut the desired gene are used to cut the vector DNA (usually the plasmid from a bacterium). This will produce ‘sticky ends’ complementary to the ends of the desired gene.

Question 6

how do the cut vector and the cut gene of interest get joined back up together

Answer 6

Mix both the cut vector DNA and the gene with DNA ligase, an enzyme that will anneal (join) the two molecules together.

Question 7

what happens once the cut gene and the cut vector are joined back together with DNA ligase

Answer 7

Mix recombinant plasmids with bacteria (E.coli). Apply temporary heat or electric shock. (This opens up the pores in the cell surface membrane of the bacterium for the plasmid to enter.)

Question 8

what

Answer 8

The transgenic bacteria are isolated and grown for mass production. The desired protein is extracted and purified before use.