1. The Gene Frequency of the CCR5
Delta 32 Allele from a Population in
Northeastern Ohio
Jacqueline Makowski, Alexis Shawver, Connor Anderson, Alexandra
Fulton, Megan Sheldon and Victoria Soewarna
Advisor: Dr. Harry Kestler

2. HIV Entry

3. Delta 32

Deletion of 32 base pairs in the CCR5 sequence

Stemmed from Eastern Europe during the time of the
Black Plague

Black Plague lasted over 500 years

This gave the human body a chance to mutate and
adapt to the plague

Most commonly found in Caucasians

Has not been studied in other races

4. Procedure
2. Determine what primers would
1. Map Primers for CCR5 gene
visually indicate Delta 32
3. Develop protocol for DNA 4. Test subject approval ( & College
sample collection Internal Review Board approval)
5. Presenting study to test subject 6. Collect samples (labeled randomly)
7. Extract DNA, PCR, Run a gel 8. Repeat process with improvements
• Analyze and Record Data • Fix mistakes (contamination, etc.)
9. Continue to test and retest
samples 10. Analyze results

5. CCr5 Sequence map
CCR5 Sequence Around Delta32 Mutation
Primers F7 & B8 will generate a 140 base pair PCR product for wild-type and a 108
base pair product Δ32 allele.
PCR conditions: (5 minutes at 95° (once) 1 minute at 95° (thirty times) 1 minute at
60°2 minutes at 72°10 minutes at 72°,then, stops at 4°) using Pure Taq Ready to
go PCR beads.

6. The effects of Δ32

Amino acids come in groups of three called codons
 GCA ATC GTA

32 is not divisible by three

Everything after the 32 base pair deletion mutation
consequently changes

For example:
 THE CAT RAN
 Remove the “C” in “CAT”

7. Protocol
1. Participants were given 2. Inside of cheek swabbed
a sterile swab for one minute
3. Swab placed in a tube
4. Tubes were randomly
containing a lysing agent to
labeled
extract
6. PCR samples were
5. PCR was performed on
analyzed by agarose gel
the samples
electrophoresis

8. +/ Δ 32
+/ Δ 32
Agarose gel
+/ Δ 32

9. Results
•
45 out of the 50 samples are homozygous wild-type
•
Normal progressor
•
5 out of the 50 samples are heterozygous for Δ32
•
Long term non-progressor
•
0 out of the 50 samples are homozygous for Δ32
•
Less likely to contract HIV

10. Conclusion
•
Five out of fifty people were found to be
heterozygous for the delta 32 deletion
mutation
•
Five alleles out of one hundred alleles or a
gene frequency of 5% was found within the
Early College Population

11. Future Plans
•
Continue testing for the Δ32 deletion
mutation at LCCC
•
Clone the Δ32 mutation into a plasmid vector
to transform hematopoietic stem cells
•
This could be used for gene therapy to
potentially cause all new blood cells to be
Δ32.