Analysis of Delta32
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This document summarizes a student research project that analyzed the frequency of the CCR5 Delta 32 allele in a population in Northeastern Ohio. The students mapped the CCR5 gene, developed a DNA collection and analysis protocol, and tested 50 samples. Their results found that 5 out of the 50 samples were heterozygous for the Delta 32 allele, indicating a gene frequency of 5% in the population. The students propose continuing their research and exploring using the Delta 32 mutation for potential gene therapy applications.
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Nucleic acids are macromolecules found in all living cells that carry genetic information. Friedrich Miescher first isolated nucleic acids from white blood cells in 1869. There are two main types of nucleic acids: DNA and RNA. DNA is the genetic material found in the nuclei of cells and in organelles like mitochondria. It has a double-helix structure formed by pairing of nitrogenous bases. RNA is also found in cells and is involved in protein synthesis. The flow of genetic information goes from DNA to RNA to protein, as described by the central dogma of molecular biology.
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Dr. Manish Jain
Scientist
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DNA is the genetic material present in all living cells. It is a double-stranded molecule composed of deoxyribose, phosphate groups, and nitrogenous bases. DNA replicates semi-conservatively to produce two identical copies of itself during cell division. It carries genetic information that is transcribed into mRNA which is then translated into proteins.
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This is a classical paper which I have selected for the explanation of the genes upregulated in Teleosts.
Chapter 6. Molecular basis of inheritance.
Nucleic acids like DNA and RNA are the genetic material found in living cells. DNA carries genetic information from one generation to the next and is made up of deoxyribose, phosphate groups, and nitrogenous bases. DNA replication is semi-conservative and produces two identical DNA molecules, each with one old and one new strand. Transcription produces mRNA from a DNA template, and translation reads mRNA to produce proteins according to the central dogma of biology.
Class9 DNA technology in secondary school
Biotechnology is the use of an organism, or a component of an organism or other biological system, to make a product or process.
Many forms of modern biotechnology rely on DNA technology.
DNA technology is the sequencing, analysis, and cutting-and-pasting of DNA.
Common forms of DNA technology include DNA sequencing, polymerase chain reaction, DNA cloning, and gel electrophoresis.
Biotechnology inventions can raise new practical concerns and ethical questions that must be addressed with informed input from all of society.
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Analysis of Delta32
- 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.
Editor's Notes
- J and A
- J
- J
- A
- A
- A
- J
- J
- A
- A
- J