Lecture 7

Question 1

How does targeted gene knockouts in yeast work

Answer 1

• Incorporate target yeast gene sequence into primers and use primers to amplify resistance gene.
• Apply as disruption construct to a diploid cell so during replication they align.
• Washing with antibiotic shows the resistant cells.

Question 2

What is RNA interference

Answer 2

dsRNA ingibits gene function by base pairing with target mRNA leading to the degredation or silencing of mRNA.

Question 3

How do you produce dsRNA

Answer 3

Sense and anti-sense RNA molecules produced in the test tube. Heated and cooled together to allow base pairing between the two.

Question 4

What is the process of transgenesis

Answer 4

1) Plasmid carrying donor gene
2) inject into the nucleus of a fertilised egg
3) Mouse gives birth and DNA is extracted as a sample.
4)Extra DNA sample and hybridise with probe for plasmid
6) Mate mice to produce a stable line expressing that gene.

Question 5

How do you use embryonic stem cells to generate a transgenic mouse

Answer 5

1) Take fertilised egg and Blastocyst
2) Remove stem cells from Blastocyst
Cells grown and cultured in tissue culture facility.
3)Gene transfer directly into mouse via blastocyst