You discover two of the three zebrafish genes for yellow pigment production are

Question

You discover two of the three zebrafish genes for yellow pigment production are expressed in your frog. You decide to look for mutations in these genes by amplifying each gene by PCR and sequencing the PCR products by Sanger sequencing to determine if any of these genes are mutant in your blue frog

a. You sequence the DNA and obtain the result below:


b. What is the chemical basis that requires the sequencing gel on the left to use four lanes and the colored image on the right to use only one?

c. Write the sequence displayed on the sequencing gel above in the 5’-3’ direction.

d. Write the corresponding sequence from the TEMPLATE strand of the DNA molecule in the frog.

e. Nucleotide changes in the sequence of a gene can lead to a change in the genetic code. These mutation include synonymous/silent, misssense/nonsynonymous and/or nonsense mutations. Which of these mutations might you expect to find in the mutant gene of your blue frog? Explain why.

Explanation / Answer

b. The short answer is that left is radioactive in exposure while the right uses fluorescent tags. c. ATG is the start codon (at the top) you just have to go down from there d. Reverse the series (start from bottom) e. Not silent, otherwise it would be green. In missense mutations, many times the gene expressed has a certain degree of functionality. Nonsense mutations would work here because the yellow appears completely nonfunctional.

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