Title:

Assessing the impact of UV irradiation on wild type E. coli and mutant E. coli deficient in
nucleotide excision repair

Introduction:
DNA controls the design of all life on earth. As it is the blueprint for our cells to do its daily
functions, various mechanisms have evolved to prevent faulty DNA from propagating and
causing havoc within our bodies. Two of such ways to fix DNA is photoreactivation repair
and nucleotide excision repair which both repair when one strand of the DNA molecule if it
has been damaged, normally be UV radiation. Photoreactivation repair does this by directly
using enzymes to find the exact right nucleotides to replace while nucleotide excision pair,
the more reliable method cut out a 24 long nucleotide strand and fully replaces it using DNA
polymerase and DNA ligase.

E. coli, a gram negative bacteria, has in its genome a lac operon, which controls whether
lactase, an enzyme which breaks down a sugar lactose, is produced. The experiment used two
strains of E. coli to measure the effect of UV irradiation on the survival of the cells. The
experiment measured the survival of the strains of E. coli based on how many E. coli colonies
were left in the petri dishes after they were incubated. The E. coli was diluted various times
to prevent too many colonies from forming and making counting the number of colonies
feasible to do. The number of colonies on the plate aimed to have between 30-300 colonies.
The results of this will then be further analysed to find the kill count for both strains.

These two samples were wild type and mutant DNA repair deficient E. coli in the two
mechanisms outlined earlier which were both irradiated in petri dishes at varying times. The
mutant E. coli had a mutation in the part which codes nucleotide excision repair and
photoreactivation, making it much more susceptible towards further mutations under UV
irradiation. After they were irradiated E. coli strains were incubated.

This experiment increases our understanding of our knowledge of DNA repair pathways
which can help various medical fields such as cancer research and gene editing for genetic
disorders. However the study of DNA repair mechanisms has much broader applications than
just medical research and can help us study microorganism environments and UV radiation’s
effect on these populations.

Aim:
To test different susceptibilities of wild type and mutant E. coli strains in UV irradiation,
assessing the effect of nucleotide excision repair and photoreactivation enzymes in DNA
damage.

Results:
Table 1: Wild Type E. coli
Number of Colonies Dilution Factor Number of cells /ml
 on plate

0 sec UV 0 10^-5 0

10 sec UV 15 10^-3 1.5 x 10^5

20 sec UV 7 10^-3 7 x 10^4

40 sec UV 78 10^-2 1.6 x 10^4

Table 2: DNA repair-deficient E. coli

Number of Colonies Dilution Factor Number of cells/ml
on plate

0 sec UV 416 10^-5 4.16 x 10^6

1 sec UV 624 10^-3 6.24 x 10^4

2 sec UV 2048 10^-2 2.05 x 10^3

4 sec UV 2592 10^-2 2.60 x 10^3

Number of Cells/ml = Number of Colonies x 1/ dilution factor x 10
Number of Cells/ml for 0 sec UV DNA repair-deficient E. coli = (416 x 1) / (10^-5 x 10) =
4.16 x 10^6
 Graph 1: Semi log graph of Kill Curve of Wild Type of DNA repair deficient E. coli to UV
light

Graph 2: Semi log graph of Kill curve of DNA deficient E. coli exposed to UV light