Cloning Flashcards
What is the first step in cloning cells?
Isolating the genomic DNA and the chosen vectors.
The plasmid is engineered to carry what?
ampR and lacZ
What is the function of ampR?
Makes E. Coli cells that are resistant to the antibiotic ampicillin
What is the function of LacZ?
It encodes B-galactosidase
What is the function of B-galactosidase?
It hydrolyzes the sugar lactose and X-gal
What does the plasmid contain?
One copy of the restriction site
What does the restriction enzyme do?
It cuts the plasmid and DNA
How is base pairing allowed between complementary sticky ends?
From the mixing of the Fragments
The addition of DNA ligase does what?
It covalently bonds the sugar phosphate backbones of the fragments
Some of the resulting recombinant plasmids contain what?
DNA fragments
One of the three recombinant plasmids carry what?
The B-globin gene
One of the three carrying the B-globin gene producing what?
Plasmids containing DNA fragments, a combination of two plasmids, or a rejoined, nonrecombinant version of the original plasmid.
What is added to bacteria?
The DNA mixture.
What is special about the bacteria?
It has a mutation in the lacZ gene.
What does the mutation cause?
It causes the the bacteria to be unable to hydrolyze lactose.
