Lecture 27

Question 1

In a test tube, how many bases can you make up?

Answer 1

150-200 bases.

Question 2

Some pUC vectors have what from bacteriophage (F1)?

Answer 2

Ori. (200-300 bp long)

Question 3

A cell with pUC plasmid and F1 phage will result in what?

Answer 3

MANY copies of that ssDNA. Always makes the same strand.

Question 4

If you take an oligonucleotide and hybridise it to ssDNA, how much will be mutant?

Answer 4

Half.

Question 5

What is NHEJ?

Answer 5

Non-homologous end joining. Cells repair dsDNA breaks. It’s more frequent than homologous recombination.

Question 6

What does Kanamycin (antibiotic) target?

Answer 6

Chloroplast and bacterial ribosomes (stops protein production).

Question 7

What does Geneticin target?

Answer 7

Cytoplasmic ribosomes.

Question 8

What does the E.coli kanamycin-resistance gene do?

Answer 8

Inactivates antibiotic via phosphorylation (kinase reaction). Good selectable marker.

Question 9

SV40 late promoter and SV40 polyA are found where?

Answer 9

In animals. It is a tumor producing gene.

Question 10

CaMV 35s promoter and CaMV polyA are found where?

Answer 10

In plants. Cauliflower Mosaic Virus.

Question 11

What is a reporter gene?

Answer 11

Monitors the level of expression of the promoter. E.g. lac, gus, luc, gfp.

Question 12

Where can you use these reporter genes? Lac, gus, luc and gfp.

Answer 12

Lac - not in plants.
gus - only in plants.
luc - firefly luciferase (most sensitive)
gfp - green fluoro protein (non-destructive)