Genetic engineering part 2 Flashcards
1
Q
What is transformation?
A
- transferring the plasmid with the recombinant DNA into the host cell
2
Q
WHat is the calcium method of transformation?
A
- culture the bacterial cells and plasmids in a calcium0rich soln and increase temp
- causes bacterial membrane to become permeable and the plasmids can enter
3
Q
What is electroporation?
A
- another method of transformation
- small current applied to bacteria
- makes membranes very porous
- plasmids move into cells
4
Q
How else can electroporation be used?
A
- to get DNA directly into eukaryotic cells
- the new DNA will pass through the cell membrane and the nuclear membrane to fuse w nuclear DNA
- less effective
5
Q
Why does current neeed to be carefully controlled?
A
- membrane can be permanetly damaged or destroyed
- in turns, may destroy whole cell
6
Q
What is electrofusion?
A
- another way of producing genetically modified cells
- electric current applied to membrane of two different cells
- this fuses the cell and nuclear membranes of the two different cells togetheer to form a hybrid or polypoid cells
- contains DNA from both
- used succesfully to produce GM plants
7
Q
How is electrofusion used in animal cells?
A
- do not fuse as effectively as plant cells
- mammalian polypoid cells do not survive in the body
- although, it is important in the production of monoclonal antiboides
8
Q
What is a monoclonal antibody?
A
- produced by a combination of a cell producing one single antibody witha tumour cell, which means it divides rapidly in culture
- now used to identify pathogen in animals and plants, and in treatment of diseases, some forms of cancer
9
Q
Which organisms are easiest to genetically modify?
A
- GM easier with prokaryotes
- plants easier to work with than animals
10
Q
Why are prokaryotes genetically modified?
A
- bacteria and other microorganisms have been GM to produce many different substances
- e.g. insulin and human growth hormones, antibiotics, pure vaccines
11
Q
How are plants genetically modified?
A
- can be modified using Agrobacterium tumefaciens, a bacterium that causes tumours in healthy plants
- desired gene (e.g. herbicide/drought resistance or pesticide production) placed in plasmid of A. tumefaciens along w marker gene
- then carried directly into the plant cell DNA
- trnasgenic plant forms a callus , which can be grown into a new plant
- electrofusion
- use same or different species
- callus fromaation then clones
12
Q
Why is it more difficult to modify animals? Why is it important?
A
- membranes more difficult to manipulate
- produce medically important proteins, e.g treatment for Huntington’s disease
