CellSig3 - 13

Question 1

What does endogenous refer to?

Answer 1

Naturally occurring genes, RNA and proteins in cells or organisms

Question 2

What does exogenous refer to?

Answer 2

Experimental manipulation of genes, RNA or proteins added to cells or organisms

Question 3

What sort of systems allow real-time measurement?

Answer 3

Electrophysiology, GFP-tags, biosensors

Question 4

What sort of systems require fixing before measurement?

Answer 4

Immunohistochemistry and biochemistry

Question 5

List three direct labelling techniques

Answer 5

Radioactive, photo/chemically reactive, GFP

Question 6

List the main indirect labelling technique

Answer 6

Immunological - a/b labelling or enzymes etc

Question 7

Outline fusion protein incorporation

Answer 7

Gene for protein of interest has DNA encoding epitope/GFP tag inserted, then introduced into cell; natural incorporation leads to purification, visualisation or immunolocalisation

Question 8

Outline immunocytochemistry

Answer 8

Epitope tag incorporated into protein of interest, primary a/b attaches to this tag, then marker-coupled secondaries attach to primary, amplifying response

Question 9

What protein-protein interaction methods can be used?

Answer 9

Co-immunoprecipitation, GST pull-downs, yeast two-hybrid, FRET

Question 10

What does FRET stand for?

Answer 10

Fluorescence resonance energy transfer

Question 11

What in vitro methos are there for studying transcriptional regulation?

Answer 11

Reporter constructs (lacZ, CAT, luciferase), RNAse protection

Question 12

What whole genome approaches are there to studying transcriptional regulation?

Answer 12

Microarrays