April 30, 2013

Sample problems:
1. A conjugation between his+ pyr+ lys+ arg+ str sensitive cells and his- pyr- lys- arg- str
resistant cells is allowed to continue for 30 minutes. At this time the mating is stopped
and the genotypes of the recombinants are determined. The results appear below. What
is the first gene to enter and what is the probable gene order?
_________________________________________
Genotype
No of colonies
_________________________________________
his+
500
+
pyr
100
lys+
350
+
arg
350
________________________________________
2. A new mutant was isolated that is unable to use acetate as a carbon source (ace). To
determine where the mutation maps, it was mated with the four different ace+ Hfr donor
strains. For simplicity all 4 Hfr strains are shown on one E. coli chromosome below.
Exconjugants were selected for growth on acetate as sole carbon source [Arrowheads
indicate the location and direction of transfer from each Hfr strain.]

Given the results in the following table, where does the ace mutation map? [Indicate the
map position in minutes.]
Donor Strain
Hfr 1
Hfr 2
Hfr 3
Hfr 4

Ace+ colonies
1000
5
1000
80

3. Glucose can be catabolized as shown below. The enzymes responsible for each
reaction is indicated by letters over the arrows:
A

Glucose (G)

glucose 6phosphate
(G6P)

D
glyceraldehyde 3-phosphate (G3P) (G3P)

fructose 6-phosphate (F6P)

C
fructose 1,6-phosphate (F1,6P)

You have 4 strains, each with a mutation in a different enzyme in the pathway. You
supplement the strains with one of the products in the pathway (i.e., you provide that
product in the medium) to see which strains can produce G3P with the supplement (+
indicates can produce G3P, - indicates cannot produce G3P). You find the following:

Strain
1
2
3
4

Media Supplement
G
G6P F6P F1,6P

+
+

+
+
+

G3P
+
+
+
+

Which strains have mutations in enzymes A and C?

4. A new proline auxotrophic mutant (pro-53) was isolated. By constructing partial
diploid strains, a complementation table was constructed as shown below. One copy of
the genes is shown on the top of the table and the other copy of the genes is shown
down one side of the table. Only the mutant genes are shown in the table. "-" indicates
failure to complement and "+" indicates complementation. What gene does the pro-53
mutation affect?
proB
proA
proC
pro-53

proB
+
+
+

proA
+
+
-

proC
+
+
+

5. Phage P22 was grown on the donor cells shown below. Using these phage lysates,
transductions were conducted with Salmonella typhimurium strains with mutations in the
metA, aceA, and iclR genes. Given the results shown in the table below, draw a genetic
linkage map showing the cotransduction frequency and the predicted order of the metA,
aceA, and iclR genes. Show your calculations. [Note that Tn10 indicates an insertion
mutation with a tetracycline (Tet) resistant phenotype. Genetic mapping of Tn10 is
essentially like any other gene.]

Selected
phenotype

Recombinants

Number
obtained

metA::Tn10 iclR- metA+ iclR+

TetR

IclRIclR+

60
40

aceA+ iclR-

aceA- iclR+

Ace+

IclRIclR+

95
5

metA::Tn10
aceA+

metA+ aceA-

TetR

AceAce+

10
90

Donor

Recipient
cells

6. Phage DNA that is r+ m+ tu+ is used to transduce a strain that is r- m- tu-. The numbers
of each transductant genotype appear below. What can be said about the relative
positions of the genes?
Genotype
r+ m+ tu+
r+ m+ tur- m- tu+
r- m- tur+ m- tu+
r+ m- tur- m+ tu+
r- m+ tuTOTAL

Number
1243
322
284
1158
175
54
58
160
3454