Plasmid Recombination

Question 1

Competent cells

Answer 1

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

Question 2

Vectors

Answer 2

-plasmids are designed to transfer foreign genetic material into a cell
-plasmids with high copy number will have many copies in a cell and therefore will create multiple products
EX. Insulin

Question 3

Cloned genes

Answer 3

Identical multiple copies of a gene

Desired DNA sequence can be joined with bacterial plasmids to form recombinant DNA

Question 4

Steps to clone a gene

Answer 4

1. DNA segment of interest is cut using a restriction enzyme
2. bacterial plasmid that is antibiotic resistant is cut with the same restriction enzyme
3. Dna and plasmid join at sticky ends to form recombinant DNA
4. Bacterial cells are transformed, the cells are than plated in a medium containing anti biotic
5. Resistant bacteria proliferated on medium, as bacterial divide recombinant plasmids replicate amplifying the piece of DNA inserted into the plasmid

Question 5

Restriction maps

Answer 5

• diagram showing the relative location of all the known restriction enzyme recognition sites on a particular plasmid and the distance between those sites
• use these maps to determine which restriction enzyme they will use to cut the plasmid

Question 6

Plasmids

Answer 6

• Small circular pieces of DNA found in bacteria
• Replicate independently of chromosomal DNA
• often have genes for proteins that provide protection from antibiotics