A blotting method known as Western blotting can be used to detect proteins that
ID: 66160 • Letter: A
Question
A blotting method known as Western blotting can be used to detect proteins that are translated from a particular mRNA. This techniques first separates proteins be size using electrophoresis. then detects specific proteins through staining (antibody directed). Suppose a researcher was interested in the effects of mutations on the expression of a gene that encodes protein X. This protein is expressed in the skin and contains 572 amino acids with a molecular mass of 68 kDa. Make a drawing that shows the expected results of a Western blot for this protein using skin cells that were obtained from the following individuals Box indicates the "ell. Indicate lane (top) and sizes (right side) of the image (12). Normal Individual an individual who is homozygous for a deletion, which promoter for this gene an individual who is heterozygous in which one gene is normal -other has a mutation that introduces an early stop codon at codon an individual w ho is homozygous for a mutation that introduces an early stop codon at codon 100. an individual who is homozygous for a mutation that than from a valine codon into a leucine codon. an indidual who is homozygous for a mutation within tin nhmu region blocking transcription factor binding.Explanation / Answer
Lane 1
Lane 2
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Lane 6
68 kD band
68 kD band
68 kD
No band
50 kD
No band
12 kD
Average molecular weight of one amino acid = 68,000 Da / 572 amino acid = 119.0 kDa
1 codon = 1 amino acid
1 codon = 3 bases
In absence of promoter, RNA polymerase can't bind and thus no transcription, no protein.
Blocking of transcription factor, does not allow recruitment of RNA polymerase to promoter region, thus no transcription, no protein.
Substitution of one amino acid by other will not have any affect on molecular mass of protein.
Introduction of stop codon early in mRNA sequence forms truncated protein.
Lane 1
Lane 2
Lane 3
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Lane 5
Lane 6
68 kD band
68 kD band
68 kD
No band
50 kD
No band
12 kD
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