Molecular Genetics

Question 1

What is the first transformation step require when cloning cells to express a specific enzyme?

Answer 1

Must transform the ‘expression cassette’ containing the gene from the enzyme into a suitable vector plasmid

Question 2

What makes the plasmid vector suitable?

Answer 2

Contains a selectable marker and an inducible promoter

Question 3

What is the purpose of ligating the cloned plasmid into E.coli?

Answer 3

To grow up the levels of plasmid and extract some DNA to ensure you have the correct plasmid - identified by PCR

Question 4

How do you linearise the plasmid to transform it into the yeast cells?

Answer 4

Using Nru1

Question 5

What induces the expression of a gene downstream of a MOX promoter?

Answer 5

Methanol

Question 6

What factor is key in quantifying the amount of DNA from a gel analysis?

Answer 6

The amount of DNA you have personally loaded in comparison to the standard gel