Selection of Mud1 Lysogens
Dilution of Lysate 
Number of Colonies 
White 

Red 

Total 

% red colonies 
SL 
J&I 
A 
SL 
J&I 
A 
SL 
J&I 
A 
SL 
J&I 
A 
10^{1} 
0 
45 
TNTC 
0 
5 
15 
0 
50 
TNTC 
0 
10% 
0 
10^{2} 
185 
0 
39 
15 
0 
5 
200 
0 
44 
7.5% 
0 
11.3% 
10^{3} 
0 
0 
0 
0 
0 
3 
0 
0 
3 
0 
0 
100% 
Uninfected 
0 
0 
0 
0 
0 
0 
0 
0 
0 
0 
0 
0 
SueLynn, Joel & Isleen’s
data provided by Mr.Sandoz
Joel and Isleen did not get any colonies because the MC4100 was not infected
by the lysate. This is why data was provided to them.
In order to select the Mud1 lysogens (above data) the plates used are Lactose MacConkey
with ampicillin. The MacConkey also has an acid base indicator and colonies that metabolize the sugar will produce
acid and turn red. In our data specifically, some of the colonies are red because Mud1 inserted next to an active promoter
in the MC4100 genome. This allows the lactose structural genes to be transcribed by the host promoter and continue to be translated
and use these protein products to metabolize the lactose.
The white
colonies are lysogenic for MuD1 and not MuC^{ts} because only MuD1 contains the bla genes which have the ability to
make the cell resistant to ampicillin.
The average
frequency of lysogeny to ampicillin resistance is 1.155 x 10^{4} Amp^{r}/pFu.
The frequency
to Ampicillin resistance assuming the Viable Cell Count of the MC4100 is 2 X 10^{9} cfu/ml is Less Than 5 x 10^{9}
mutations/vc.
Calculations:
Frequency
of lysogeny to amp^{r}:
SueLynn’s
Data
200 / .1 mL= 2000 lysogens/mL
4 x 10^{9 }pFu/mL x 10^{2} x 1/3 = 1.33 x 10^{7}
2000/ 1.33 x 10^{7}= 1.5 x 10^{4} Amp^{r}/pFu
Joel
& Isleens’s Data
50 / .1 mL= 500 lysogens/mL
2.73 x 10^{9} pFu/ml x 10^{1} x 1/3 = 9.1 x 10^{7}
500/9.1 x 10^{7}= 5.5 x 10^{6} Amp^{r}/pFu
Alex’s
Data
44/0.1mL = 440
6.9x10^{8}
x 10^{2} x 1/3 = 2.3x10^{6 }
440/2.3x10^{6}
= 1.91 x 10^{4} Amp^{r}/pFu
Average
Frequency of lysogeny to amp^{r}:
(1.5 x 10^{4} Amp^{r}/pFu + 5.5 x 10^{6}
Amp^{r}/pFu + 1.91 x 10^{4} Amp^{r}/pFu)/3 = 1.155 x 10^{4} Amp^{r}/pFu
Mutation
Frequency:
Everyone’s
Data
1/.1 mL= 10 cFu/mL
10/ 2 x 10^{9}= Less Than 5 x 10^{9} mutations/vc
Selection of the Ara^{r} Lysogens
Dilution of lysate 
Number and Description of the Colonies 
SueLynn 

Joel & Isleen 

Alex 
Undiluted 
10 W 
1 R 
12 W 
4 R 
0 W 
0 R 
10^{1} 
0 W 
0 R 
0 W 
0 R 
5 W 
20 R 
10^{2} 
0 W 
0 R 
0 W 
0 R 
0 W 
0 R 
Uninfected Mc4100 
150 W 
50 R 
141 w 
32 R 
0 W 
0 R 
All data provided by Mr.Sandoz
Theoretically
there should be colonies on the uninfected MC4100 plate but the probable source of no colonies here is human error. The lysate
may have accidently been plated. For accurate calculation purposes we will only use SueLynn, Joel and Isleen’s data
to calculate the average concentration of Ara^{r} mutations and Ara^{+} mutations.
The average frequency of lysogeny to Arabinose resistance is 7.92 x 10^{7}
Ara^{r}/pFu.
The ratio
of Arar lysogens/pfu: Ampr lysogens/pfu is:

SueLynn 
Joel & Isleen 
Alex 
Ratio 
1
: .0005 
1
: .024 
1
: .011 
The average
concentration of Ara^{r} mutations is (1500 + 1410)/2= 1455 cfu/mL
The average concentration of Ara^{+ }mutations is (500 + 320)/2= 410 cfu/mL
The average mutation frequency for each of the above is:
Ara^{r }mutations: (7.5 x 10^{7} + 7.05 x 10^{7})/2= 7.28 x 10^{7} cFu/mL
Ara^{+ }mutations: (2.5 x 10^{7} + 1.6 x 10^{7})/2=
2.05 x 10^{7} cFu/mL
The most
likely reason that these frequencies are high is because Ara^{r} was induced by the Mud1 insertion and was not a spontaneous
mutation. The Mud1 could insert anywhere in the arabinose operon, in which multiple
genes are involved, which increases the odds of causing a change in phenotype. There are also multiple chances for mutation
because there are multiple generations before ribulose5phosphate kills the cell.
Calculations:
SueLynn’s
Data
 10 W colonies/.1mL = 100 cFu/mL
100/
(4.0 x 10^{9})(1)(1/3) = 7.5 x 10^{8}A Ara^{r}/pFu
 7.5 x 10^{8} Ara^{r}/pFu
: 1.5 x 10^{4} Amp^{r}/pFu
1
: .0005
 150 cFu/.1mL= 1500 cFu/mL
 50 cFu/.1 ml= 500 cFu/mL
 1500/2 x 10^{9}= 7.5 x 10^{7} cFu/mL
500/2
x 10^{9}= 2.5 x 10^{7} cFu/mL
Joel
& Isleen Data
 12 W colonies/.1mL = 120 cFu/mL
120/
(2.73 x 10^{9})(1)(1/3) = 1.31 x 10^{7}A Ara^{r}/pFu
 1.31 x 10^{7}A Ara^{r}/pFu : 5.5 x 10^{6} Amp^{r}/pFu
1
: .024
 141 cFu/.1mL= 1410 cFu/mL
 32 cFu/.1 ml= 320 cFu/mL
 1410/2 x 10^{9}= 7.05 x 10^{7 }cFu/mL
320/2
x 10^{9}= 1.6 x 10^{7} cFu/mL
Alex’s
Data
 5 W colonies/.1mL = 50 cFu/mL
50/
(6.9 x 10^{8})(10^{1})(1/3) = 2.17 x 10^{6 }Ara^{r}/pFu
 2.17 x 10^{6 }Ara^{r}/pFu
: 1.91 x 10^{4} Ampr/pFu
1
: .011
Average frequency of lysogeny to Arabinose resistance:
(7.5
x 10^{8}A Ara^{r}/pFu + 1.31 x 10^{7}A Ara^{r}/pFu + 2.17 x 10^{6 }Ara^{r}/pFu)/3=
7.92 x 10^{7} Ara^{r}/pFu