Plasmid Recombination

Question 1

Plasmid recombination

Competent cells?

Answer 1

Cells that are able to take up foreign (plasmid) DNA

Question 2

Plasmid recombination

Vectors?

Answer 2

-plasmids are designed to transfer foreign genetic material into cell
-plasmids with high copy numbers will have many copies and create multiple products
Ex. A plasmid with high copy number engineered to produce insulin, will produce more than a plasmid with low copy number

Question 3

Plasmid recombination

Cloned genes?

Answer 3

• identical multiple copies of a gene

- desired DNA sequences can be joined with bacterial plasmids to form recombinant DNA

Question 4

Plasmid recombination

Steps to clone a gene? 5?

Answer 4

1. DNA segment of interest is cut using restriction enzyme
2. Bacterial plasmid that is resistant to antibiotics is cut with same restriction enzyme
3. DNA and plasmid join at sticky ends to form recombinant DNA
4. Bacterial cells are transformed, then played on medium containing antibiotic
5. Resistant bacteria proliferate on medium. As bacteria divided recombinant DNA replicate amplifying the inserted DNA piece

Question 5

Plasmid recombination

Restriction maps?

Answer 5

• a diagram that shows the relative location of all restriction enzyme recognition sites on a plasmid and the distance between those sites
• scientist use these maps to find out which restriction enzyme they will use to cut the plasmid

Question 6

Plasmid recombination

Plasmids?

Answer 6

• small circular price of DNA found in bacteria
• replicate independently on chromosome in DNA
• often have genes for protein, provide protection from antibiotic