genetic engineering Flashcards
1
Q
state the 5 steps of DNA technology (in vivo cloning)
A
- create DNA fragment
- insert DNA into a vector
- transform host cell with vector
- identify transformed cell
- grow host cell (clone)
2
Q
how is DNA amplified?
A
- using PCR
- cDNA into thermal cycler
3
Q
summarise how bacteria is used to produce human insulin
A
- mRNA extracted from cells and treated with reverse transcriptase to make complementary DNA/plasmid obtained from bacteria & cut with restriction enzyme
- plasmid and cDNA fuse using DNA ligase
- recombinant plasmid introduced into host cells
- bacteria multiply in a fermenter and produce insulin
- separation & purification of human insulin can be used by diabetic patients
4
Q
what are the advantages of using bacteria in this process?
A
- multiply quickly
- DNA is easily accessible (has free/plasmid DNA in cytoplasm)
5
Q
describe how a plasmid is cut
A
- both strands of DNA cut
- using a specific restriction enzyme
- in a particular section
- creates sticky ends
6
Q
how can the same sticky ends be created?
A
use same restriction enzyme
- will cut at same recognition site
7
Q
why are sticky ends made to be identical?
A
cut plasmid & cDNA align and the cDNA becomes part of the plasmid
8
Q
how can the plasmid be inserted into a bacterium?
A
- injected (using a vector)
- use a solvent / detergent
- increase temperature
- electric shock bacteria cell
9
Q
how can bacteria that have taken up insulin be detected?
A
inserting an indicator/antibiotic resistant bacteria when cDNA is inserted to show those that have picked up the plasmids
10
Q
A
