In Vivo Cloning Flashcards
What is in Vivo cloning
Transferring fragments to a host cell using a vector
How r we able insert gene into vector
Sticky ends
Most common type of vector
Plasmids
How are DNA fragments inserted into a plasmid
- restriction enzyme cuts plasmid
- produces sticky ends
- ligand joins DNA and plasmid
What allows DNA sticky ends to be complementary to vectors sticky ends
Gene cuts from DNA using restriction enzymes and the SAME restriction enzyme must be used to cut vector
What joins complementary bases together
Hydrogen bonds
Enzyme that joins vector and DNA
Ligase
summarise the process of inserting a DNA fragment into a vector
a plasmid is used as a vector and is cut using the same restriction enzyme as the DNA so ends are complementary , produces sticky ends. DNA ligase joins fragments and plasmid together
how do u insert a vector into a host cell
use ice cold calcium chloride solution, heat shock, doing this increases permeability of the bacterial plasma membrane
why are ca2+ added when inserting vectors into host cells
increase the permeability of the bacterial membrane to plasmids
why are bacterial cells used as the host cell
divide rapidly
why do only a small amount of bacterial cells take up the plasmid
- some plasmids self ligate and recreate the original plasmid
- DNA fragments join together forming plasmids
what occurs if cell division happens before gene is added to the vector
gametes don’t receive the gene
how do u identify host cells that carry desirable genes
marker genes
types of marker genes
- antibiotic resistance
- enzyme
- florescent
