Gene Cloning and Transformation

Question 1

Gene Cloning

Answer 1

Inserting foreign DNA into plasmids to copy DNA as bacteria replicates

Question 2

Plasmid

Answer 2

Self-replicating, closed circular piece of DNA that can act as carrier of gene cloned in bacteria –> useful as it transfers antibiotic resistance between bacteria

Question 3

Process: Creating Recombinant DNA

Answer 3

1. YFG cut from DNA using restriction enzymes like ECORI (comes from E-coli), produces sticky ends
2. Plasmid with ampicillin resistance isolated from bacteria, and cut with EcoRI to create gap
3. YFG and plasmid combined and joined with ligase

Question 4

Transformation

Answer 4

Way to get cloned plasmid into bacteria –> bacteria acts as “factory” to produce YFG (you need its ribosomes to build proteins from YFG)

Question 5

Process: Transformation and Isolating Bacteria with Plasmid

Answer 5

1. Plasmid added to bacterial cells like e-coli (transformation)
2. Some bacteria take in plasmid, some do not
3. Bacteria plate on agar (food) that contains ampicillin
4. Those that don’t contain plasmid die, and those that do live and reproduce (can mass produce YFG)