Non-genetic analysis

Question 1

How can you detect where proteins are in the cell?

Answer 1

Tagged antibodies

Question 2

How can you detect antibodies?

Answer 2

Fluorescent tags- detected via a fluorescent microscope/ photodetector
Enzyme conjugates-
alkaline phosphatase- substrate turns blue
horseradish peroxidase- substrate turns brown

Question 3

What is the purpose of a ‘antibody sandwich’?

Answer 3

Secondary antibodies- amplify the signal- because multiple secondary antibodies can bind to a primary antibody

Question 4

What is an antisense probe?

Answer 4

Antisense (synthesised from cdna) probe hybridises with complementary mRNA- blocks its translation

Question 5

What are the uses for a GFP transgenic line?

Answer 5

1. To follow the expression of a gene in the whole animal
2. To follow the sub-cellular localisation of a protein
3. To follow the behaviour of cells- marked cells can be distinguished from others