Question

You are studying disease X, a genetic disorder that arises from a trinucleotide repeat expansion mutation...

You are studying disease X, a genetic disorder that arises from a trinucleotide repeat expansion mutation which is a mutation where entire codons are multiplied during the replication process. The mutation is found in the CaM gene, that is a 450-base pair (bp) sequence. The trinucleotide repeat expansion results in the addition of 10 codons (30 bp) at nucleotide 150 (ie in the centre of the gene, away from the 5’ and 3’ end). The sequence of the normal gene is shown below.

ATGGCTGATCAGCTGACCGAAGAACAGATTGCTGAATTCAAGGAAGCCTTCTCCCTATTTGATAAAGATG GCGATGGCACCATCACAACAAAGGAACTTGGAACTGTCATGAGGTCACTGGGTCAGAACCCAACAGAAGC TGAATTGCAGGATATGATCAATGAAGTGGATGCTGATGGTAATGGCACCATTGACTTCCCCGAATTTTTG ACTATGATGGCTAGAAAAATGAAAGATACAGATAGTGAAGAAGAAATCCGTGAGGCATTCCGAGTCTTTG ACAAGGATGGCAATGGTTATATCAGTGCAGCAGAACTACGTCACGTCATGACAAACTTAGGAGAAAAACT AACAGATGAAGAAGTAGATGAAATGATCAGAGAAGCAGATATTGATGGAGACGGACAAGTCAACTATGAA GAATTCGTACAGATGATGACTGCAAAATGA

a) Design primers for the amplification of this gene

b) Design a PCR-based method to detect this disorder in patients.

c) Describe or draw your expected results for an individual with the disease vs. a healthy individual control.

d) A sample collected from a patient with similar symptoms to those suffering from Disease X has been brought to the lab. You use your PCR based test and determine that the CaM gene is the same size as that of a healthy individual. You suspect that the arises from a point mutation (a single nucleotide change that results in a defect in the protein.) Describe a how you could confirm this hypothesis.

Homework Answers

Answer #1

a) Primers for the amplification of the entire gene:

Forward primer: 5'-ATGGCTGATCAGCTGACCGAAG-3'

Reverse primer: 5'-TCATTTTGCAGTCATCATCTGTAC-3'

b) The primers must flank the region of difference between the WT and mutant.

Forward primer: 5'-CAACAGAAGCTGAATTGCAG-3'

Reverse primer: 5'-CAAAAATTCGGGGAAGTCAA-3'

c) Isolate genomic DNA from WT and mutant individuals. Perform PCR using the given primers. Run the PCR products on the gel.

WT product size = 80 bp

Mutant product size = 110 bp

d) See image

e) If the disease is not due to triplet repeat expansion, we have to sequence the entire gene for the presence of any other SNP.

Please rate the solution. Your efforts are highly appreciated. Thank you.

Know the answer?
Your Answer:

Post as a guest

Your Name:

What's your source?

Earn Coins

Coins can be redeemed for fabulous gifts.

Not the answer you're looking for?
Ask your own homework help question
Similar Questions
You are studying disease X, a genetic disorder that arises from a trinucleotide repeat expansion mutation...
You are studying disease X, a genetic disorder that arises from a trinucleotide repeat expansion mutation which is a mutation where entire codons are multiplied during the replication process. The mutation is found in the CaM gene, that is a 450-base pair (bp) sequence. The trinucleotide repeat expansion results in the addition of 10 codons (30 bp) at nucleotide 150 (ie in the centre of the gene, away from the 5’ and 3’ end). The sequence of the normal gene...
I am hypothetically looking at a genetic disease arising from a trinucleotide repeat expansion (30 base...
I am hypothetically looking at a genetic disease arising from a trinucleotide repeat expansion (30 base pairs in total) after the 150'th nucleotide of a 450 nucleotide gene. The question asks you to design a PCR-based method to detect this disorder. We learned about qPCR and gel electrophoresis. GE: I was thinking that after PCR you could use gel electrophoresis to distinguish the size difference between the amplified product using the same primers. But at the same time I don't...
You will need to read the following article “In Iceland’s DNA Clues to Disease” (in your...
You will need to read the following article “In Iceland’s DNA Clues to Disease” (in your problem sets folder in CANVAS). The article describes the results obtained by researchers at deCODE, a genetics firm in Iceland. Their goal was to harness Iceland’s unique homogenous population with low genetic diversity coupled with excellent geneology records to uncover rare genetic mutations that could lead to disease. The research article (known as a primary source) for the information above was published in the...
"The orange locus in cats is located on the X chromosome. In females, XoXo results in...
"The orange locus in cats is located on the X chromosome. In females, XoXo results in orange cats and X+X+ results in black cats When a female is heterozygous XoX+, they express both colors in patches due to a phenomenon called X-inactivation in which one X in every cell is inactivated. These females are called calico or tortoiseshell, depending on the presence of white spots. Male cats are either orange (XoY) or black (X+Y). A cross between a black female...
The picture shown below shows variation among three individuals with respect to 4 nucleotides - AGAT....
The picture shown below shows variation among three individuals with respect to 4 nucleotides - AGAT. What do you think what type of variation is this? AGAT different repeat numbers in different individuals .png Minisatellite Single nucleotide polymorphism Short Tandem Repeats Which of the following statements is TRUE about DNA matching? Typical difference between the genomes of human beings and Chimpanzees is estimated to be 25 % Typical difference between the genomes of human beings and Drosophila is   estimated to...