The Salmon gene is GFP tagged in a human tissue culture cell line. One of the ce

Question

The Salmon gene is GFP tagged in a human tissue culture cell line. One of the cells in the line acquires a nonsense mutation in the Salmon gene.

i. Which of the below methods would allow one to identify the presence of the nonsense mutation? Select all that apply.

A. Northern blot

B. Western blot

C. RNA Seq

D. Immunoprecipitation

ii. For the above technique(s) that you selected, describe the experimental result that would be observed from them assuming there is a nonsense mutation.

iii. If you were detecting the nonsense mutation through the GFP tag, does it matter if the GFP gene was added at the 5’ end of the Salmon gene versus the 3’ end? YES or NO. Which of the statements below best describes your selection.

A. As long as the promoter is unaffected, protein will still be produced.

B. Anything downstream of the nonsense mutation will not be translated.

C. Anything downstream of the nonsense mutation will not be transcribed.

D. As long as there is no mRNA splicing occurring, exon junction complexes will not impact the process.

Explanation / Answer

1.The methods to identify the presence of nonsense mutation are: Nothern blot , Western blot and

Immunoprecipitation

2.An nonsense mutation replaces an amino acid with truncated protein

Northern western Blotting: Method which combines Northern and western blot used for identification of protein and RNA interactions

A protein population is first separated by size during gel electrophoresis followed by blotting onto a membrane

The membrane is treated with buffer containing labelled RNA proteins are able to bind RNA molecules that can be identified based on lebel(radioactive or fluorescent dye)

3.B)Anything downstream of nonsense mutation will be translated

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